ryanodine  (Tocris)

 
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  • 99
    Name:
    Ryanodine
    Description:
    Ca2 release inhibitor
    Catalog Number:
    1329
    Price:
    None
    Purity:
    ≥98% (HPLC)
    Category:
    Ryanodine Receptor Inhibitors Ryanodine Receptors Calcium Channels Ion Channels Pharmacology
    Formula:
    1H-Pyrrole-2-carboxylic acid, (3S,4R,4aR,6S,7S,8R,8aS,8bR,9S,9aS)-dodecahydro-4,6,7,8a,8b,9a-hexahydroxy-3,6a,9-trimethyl-7-(1-methylethyl)-6,9-methanobenzo[1,2]pentaleno[1,6-bc]furan-8-yl ester
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    Structured Review

    Tocris ryanodine
    Ryanodine
    Ca2 release inhibitor
    https://www.bioz.com/result/ryanodine/product/Tocris
    Average 99 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    ryanodine - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Ryanodine Receptor-Mediated Calcium Release Has a Key Role in Hippocampal LTD Induction"

    Article Title: Ryanodine Receptor-Mediated Calcium Release Has a Key Role in Hippocampal LTD Induction

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2018.00403

    LTD-induced phosphorylation of CaMKII and Synapsin I. Induction of LTD for 60 min in control (C) slices caused a significant increase in the phosphorylation levels of Synapsin I (left panels), CaMKII-α (center panels) and CaMKII-β (right panels) relative to the levels displayed by unstimulated slices (naïve). Slices pre-incubated for 1 h with 20 μM ryanodine (Rya) before applying the LTD induction protocol displayed significantly lower increments in the phosphorylation levels of Synapsin I and CaMKII-α, whereas the phosphorylation levels of CaMKII-β were not significantly different from the levels displayed by unstimulated slices. Values represent Mean ± SE ( n = 3). Statistical analysis was performed with one-way ANOVA, followed by Tukey’s post hoc test. * p
    Figure Legend Snippet: LTD-induced phosphorylation of CaMKII and Synapsin I. Induction of LTD for 60 min in control (C) slices caused a significant increase in the phosphorylation levels of Synapsin I (left panels), CaMKII-α (center panels) and CaMKII-β (right panels) relative to the levels displayed by unstimulated slices (naïve). Slices pre-incubated for 1 h with 20 μM ryanodine (Rya) before applying the LTD induction protocol displayed significantly lower increments in the phosphorylation levels of Synapsin I and CaMKII-α, whereas the phosphorylation levels of CaMKII-β were not significantly different from the levels displayed by unstimulated slices. Values represent Mean ± SE ( n = 3). Statistical analysis was performed with one-way ANOVA, followed by Tukey’s post hoc test. * p

    Techniques Used: Incubation

    Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on fEPSP rise times, half-widths and decay constants tau measured in CA3–CA1 hippocampal synapses. Left panels: slices were treated with 1 μM ryanodine. (A) fEPSP rise times vs. stimulus intensity. (B) fEPSP half-widths vs. stimulus intensity; (C) fEPSP decay constants (tau) vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine for 60 min. (D) fEPSP rise times vs. stimulus intensity; (E) fEPSP half-widths vs. stimulus intensity; (F) fEPSP decay constants (tau) vs. stimulus intensity. Right panels: slices were treated for 15 min with 1 mM caffeine. (G) fEPSP rise times vs. stimulus intensity; (H) fEPSP half-widths vs. stimulus intensity; (I) fEPSP decay constants (tau) vs. stimulus intensity. Values represent Mean ± SE; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed Mann-Whitney test (* p
    Figure Legend Snippet: Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on fEPSP rise times, half-widths and decay constants tau measured in CA3–CA1 hippocampal synapses. Left panels: slices were treated with 1 μM ryanodine. (A) fEPSP rise times vs. stimulus intensity. (B) fEPSP half-widths vs. stimulus intensity; (C) fEPSP decay constants (tau) vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine for 60 min. (D) fEPSP rise times vs. stimulus intensity; (E) fEPSP half-widths vs. stimulus intensity; (F) fEPSP decay constants (tau) vs. stimulus intensity. Right panels: slices were treated for 15 min with 1 mM caffeine. (G) fEPSP rise times vs. stimulus intensity; (H) fEPSP half-widths vs. stimulus intensity; (I) fEPSP decay constants (tau) vs. stimulus intensity. Values represent Mean ± SE; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed Mann-Whitney test (* p

    Techniques Used: MANN-WHITNEY

    Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on paired-pulse (PP) responses measured in CA3–CA1 hippocampal synapses. (A) Representative fEPSP traces showing PP responses before and after addition of 1 μM ryanodine (left panels), 20 μM ryanodine (center panels) and 1 mM caffeine (right panels). (B) Effects of 1 μM ryanodine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (D) Effects of 1 mM caffeine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (C) Effects of 20 μM ryanodine applied for 60 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. Values represent Mean ± SE; (13, 4) for ryanodine-treated slices; (12, 3) for caffeine-treated slices and (16, 4) for the control. The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used.
    Figure Legend Snippet: Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on paired-pulse (PP) responses measured in CA3–CA1 hippocampal synapses. (A) Representative fEPSP traces showing PP responses before and after addition of 1 μM ryanodine (left panels), 20 μM ryanodine (center panels) and 1 mM caffeine (right panels). (B) Effects of 1 μM ryanodine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (D) Effects of 1 mM caffeine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (C) Effects of 20 μM ryanodine applied for 60 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. Values represent Mean ± SE; (13, 4) for ryanodine-treated slices; (12, 3) for caffeine-treated slices and (16, 4) for the control. The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used.

    Techniques Used:

    Stimulatory and inhibitory ryanodine concentrations and caffeine modify the long-term depression (LTD) response. (A) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the low frequency stimulation (LFS) protocol to control hippocampal slices (14, 4) or to slices treated with 1 μM ryanodine (14, 3) or 20 μM ryanodine (13, 4). Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min after applying the LFS protocol (trace 2) to control slices, or recorded in slices treated with 1 μM or 20 μM ryanodine are shown on top of the graph. Open symbols: control slices; gray symbols: slices treated with 1 μM ryanodine; black symbols: slices treated with 20 μM ryanodine. (B) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation. (C) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the LFS protocol to control hippocampal slices (15, 3) or to slices treated with 1 mM caffeine (12, 4). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min (trace 2) after applying the LFS protocol to control slices and to slices treated with 1 mM caffeine are shown on top of the graph. Open symbols: control slices; black symbols: slices treated with 1 mM caffeine. (D) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation of control or caffeine-treated slices. Values represent Mean ± SE. Statistical significance of values was assessed by Mann-Whitney test (* p
    Figure Legend Snippet: Stimulatory and inhibitory ryanodine concentrations and caffeine modify the long-term depression (LTD) response. (A) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the low frequency stimulation (LFS) protocol to control hippocampal slices (14, 4) or to slices treated with 1 μM ryanodine (14, 3) or 20 μM ryanodine (13, 4). Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min after applying the LFS protocol (trace 2) to control slices, or recorded in slices treated with 1 μM or 20 μM ryanodine are shown on top of the graph. Open symbols: control slices; gray symbols: slices treated with 1 μM ryanodine; black symbols: slices treated with 20 μM ryanodine. (B) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation. (C) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the LFS protocol to control hippocampal slices (15, 3) or to slices treated with 1 mM caffeine (12, 4). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min (trace 2) after applying the LFS protocol to control slices and to slices treated with 1 mM caffeine are shown on top of the graph. Open symbols: control slices; black symbols: slices treated with 1 mM caffeine. (D) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation of control or caffeine-treated slices. Values represent Mean ± SE. Statistical significance of values was assessed by Mann-Whitney test (* p

    Techniques Used: MANN-WHITNEY

    Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on basal synaptic transmission in the CA3–CA1 hippocampal synapse. Left panels: slices were treated with 1 μM ryanodine; representative field excitatory postsynaptic potential (fEPSP) traces registered before and 15 min after ryanodine addition are illustrated on top of the panels. (A) Fiber volley (FV) amplitude vs. stimulus intensity; (B) amplitude vs. stimulus intensity; (C) fEPSP slopes vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine; representative fEPSP traces registered before and 60 min after ryanodine addition are illustrated on top. (D) FV amplitude vs. stimulus intensity; (E) fEPSP amplitude vs. stimulus intensity; (F) fEPSP slopes vs. stimulus intensity. Right Panels: slices were treated with 1 mM caffeine; representative fEPSP traces registered before and 15 min after caffeine addition are illustrated on top. (G) FV amplitude vs. stimulus intensity; (H) fEPSP amplitude vs. stimulus intensity; (I) fEPSP slopes vs. stimulus intensity. Values represent Mean ± SEM; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed by Mann-Whitney test ( p > 0.05 in all cases).
    Figure Legend Snippet: Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on basal synaptic transmission in the CA3–CA1 hippocampal synapse. Left panels: slices were treated with 1 μM ryanodine; representative field excitatory postsynaptic potential (fEPSP) traces registered before and 15 min after ryanodine addition are illustrated on top of the panels. (A) Fiber volley (FV) amplitude vs. stimulus intensity; (B) amplitude vs. stimulus intensity; (C) fEPSP slopes vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine; representative fEPSP traces registered before and 60 min after ryanodine addition are illustrated on top. (D) FV amplitude vs. stimulus intensity; (E) fEPSP amplitude vs. stimulus intensity; (F) fEPSP slopes vs. stimulus intensity. Right Panels: slices were treated with 1 mM caffeine; representative fEPSP traces registered before and 15 min after caffeine addition are illustrated on top. (G) FV amplitude vs. stimulus intensity; (H) fEPSP amplitude vs. stimulus intensity; (I) fEPSP slopes vs. stimulus intensity. Values represent Mean ± SEM; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed by Mann-Whitney test ( p > 0.05 in all cases).

    Techniques Used: Transmission Assay, MANN-WHITNEY

    2) Product Images from "Maternal high-altitude hypoxia and suppression of ryanodine receptor-mediated Ca2+ sparks in fetal sheep pulmonary arterial myocytes"

    Article Title: Maternal high-altitude hypoxia and suppression of ryanodine receptor-mediated Ca2+ sparks in fetal sheep pulmonary arterial myocytes

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00009.2012

    Effect of 30 mM K + on whole-cell spatial and temporal Ca 2+ signaling characteristics recorded in situ. A : baseline-subtracted fractional fluorescence (F/F o ) traces for Fluo-4 fluorescence are shown for individual pulmonary arterial myocytes from a normoxic adult sheep. Recordings were made in the absence (control) or presence of 30 mM K + (30K) with or without 10 μM ryanodine (30K RY). Bars indicate means ± SE for normoxic (open) and long-term hypoxic (solid) conditions. B, D, and F : number of myocytes with Ca 2+ responses each minute in a 1,000 μm 2 area. C, E, and G : frequency of Ca 2+ events *,§,† P
    Figure Legend Snippet: Effect of 30 mM K + on whole-cell spatial and temporal Ca 2+ signaling characteristics recorded in situ. A : baseline-subtracted fractional fluorescence (F/F o ) traces for Fluo-4 fluorescence are shown for individual pulmonary arterial myocytes from a normoxic adult sheep. Recordings were made in the absence (control) or presence of 30 mM K + (30K) with or without 10 μM ryanodine (30K RY). Bars indicate means ± SE for normoxic (open) and long-term hypoxic (solid) conditions. B, D, and F : number of myocytes with Ca 2+ responses each minute in a 1,000 μm 2 area. C, E, and G : frequency of Ca 2+ events *,§,† P

    Techniques Used: In Situ, Fluorescence

    3) Product Images from "On the mode of action of emodepside: slow effects on membrane potential and voltage-activated currents in Ascaris suum"

    Article Title: On the mode of action of emodepside: slow effects on membrane potential and voltage-activated currents in Ascaris suum

    Journal: British Journal of Pharmacology

    doi: 10.1111/j.1476-5381.2011.01428.x

    Effect of emodepside (1 µM) on membrane spiking. (A) Representative current-clamp trace showing the spiking induced by ryanodine (1 µM) and the inhibitory effect of emodepside on the spikes. (B) Bar chart (mean ± SEM) of the effects
    Figure Legend Snippet: Effect of emodepside (1 µM) on membrane spiking. (A) Representative current-clamp trace showing the spiking induced by ryanodine (1 µM) and the inhibitory effect of emodepside on the spikes. (B) Bar chart (mean ± SEM) of the effects

    Techniques Used:

    4) Product Images from "Size-dependent heterogeneity of contractile Ca2+ sensitization in rat arterial smooth muscle"

    Article Title: Size-dependent heterogeneity of contractile Ca2+ sensitization in rat arterial smooth muscle

    Journal: The Journal of Physiology

    doi: 10.1113/jphysiol.2012.241315

    Effect of treatment with 1 μ m ryanodine (rye, A ) and 1 μ m nicardipine (nic, B ) on time course of Ca 2+ rise in response to 30 μ m PE in small mesenteric artery
    Figure Legend Snippet: Effect of treatment with 1 μ m ryanodine (rye, A ) and 1 μ m nicardipine (nic, B ) on time course of Ca 2+ rise in response to 30 μ m PE in small mesenteric artery

    Techniques Used:

    Effect of 1 μ m ryanodine, 1 μ m nicardipine, and a combination of the two blockers on 30 μ m PE-induced contraction in mesenteric ( A ; n = 3–6), caudal ( B ; n = 4–6) and aortic arteries ( C ; n = 4)
    Figure Legend Snippet: Effect of 1 μ m ryanodine, 1 μ m nicardipine, and a combination of the two blockers on 30 μ m PE-induced contraction in mesenteric ( A ; n = 3–6), caudal ( B ; n = 4–6) and aortic arteries ( C ; n = 4)

    Techniques Used:

    5) Product Images from "Cerebellar Kainate Receptor-Mediated Facilitation of Glutamate Release Requires Ca2+-Calmodulin and PKA"

    Article Title: Cerebellar Kainate Receptor-Mediated Facilitation of Glutamate Release Requires Ca2+-Calmodulin and PKA

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2018.00195

    Facilitation of glutamate release mediated by presynaptic kainate receptor (KAR) activation requires an increase of Ca 2+ in the cytosol at PF-PuC synapses. (A) Time-course of KA (3 μM) effect on eEPSCs amplitude in control condition (circles) and in slices treated with philanthotoxin (squares). (B) Quantification of modulation observed in (A) . (C) Time-course of the effect of KA on eEPSCs amplitude in control slices (circles) and in thapsigargin-treated slices (squares). (D) In slices treated with thapsigargin or ryanodine, the increase of eEPSCs amplitude induced by KA is prevented. The number of slices (from two to three mice) is indicated in parenthesis at the top of each bar. Results are expressed as means ± SEM (** P
    Figure Legend Snippet: Facilitation of glutamate release mediated by presynaptic kainate receptor (KAR) activation requires an increase of Ca 2+ in the cytosol at PF-PuC synapses. (A) Time-course of KA (3 μM) effect on eEPSCs amplitude in control condition (circles) and in slices treated with philanthotoxin (squares). (B) Quantification of modulation observed in (A) . (C) Time-course of the effect of KA on eEPSCs amplitude in control slices (circles) and in thapsigargin-treated slices (squares). (D) In slices treated with thapsigargin or ryanodine, the increase of eEPSCs amplitude induced by KA is prevented. The number of slices (from two to three mice) is indicated in parenthesis at the top of each bar. Results are expressed as means ± SEM (** P

    Techniques Used: Activation Assay, Mouse Assay

    6) Product Images from "Kainate Receptor Activation Shapes Short-Term Synaptic Plasticity by Controlling Receptor Lateral Mobility at Glutamatergic Synapses"

    Article Title: Kainate Receptor Activation Shapes Short-Term Synaptic Plasticity by Controlling Receptor Lateral Mobility at Glutamatergic Synapses

    Journal: Cell Reports

    doi: 10.1016/j.celrep.2020.107735

    Desensitized LiGluK2 Receptors Are Reversibly Trapped at Glutamatergic Synapses (A) Schematic of SPT experiments: 488 nm and 380 nm light illumination was used to track receptors in the closed (blue) and desensitized (purple) states, respectively. A second 488-nm pulse induced recovery in the closed state (gray). The protocol was repeated five times every 2 min. (B) Example trajectories (yellow) of the same individual LiGluK2 receptor diffusing at synapses (Homer1c, red) in the states described in (A). Scale bar, 1 μM. (C and D) Summary of median diffusion coefficient (±IQR), immobile fraction, and MSD versus time curves of synaptic LiGluK2 (C) (n trajectories = 140, in 10 neurons from three independent cultures) and extrasynaptic LiGluK2 (D) (n trajectories = 250, in 10 neurons from three independent cultures) in the closed, desensitized, and recovery closed states. (E) Schematic of SPT experiments as in (A), in the continued presence of VGCC blockers (2-APB, D-APV, ω-conotoxin MVIIC, GYKI 53655, nifedipine, and ryanodine; black) delivered after the initial 488-nm illumination (blue). (F) Example trajectories (yellow) of an individual LiGluK2 receptor diffusing over a portion of dendrite (green) in the indicated states. Homer1c indicates synapses (red). Scale bar, 1 μM. (G and H) Summary of median diffusion coefficient, immobile fraction, and MSD versus time curve of synaptic (G) and extrasynaptic (H) LiGluK2 in different states as indicated in (E) (n trajectories : synaptic = 100; extrasyaptic = 206). Unless otherwise stated, data are presented as mean ± SEM, ∗ p
    Figure Legend Snippet: Desensitized LiGluK2 Receptors Are Reversibly Trapped at Glutamatergic Synapses (A) Schematic of SPT experiments: 488 nm and 380 nm light illumination was used to track receptors in the closed (blue) and desensitized (purple) states, respectively. A second 488-nm pulse induced recovery in the closed state (gray). The protocol was repeated five times every 2 min. (B) Example trajectories (yellow) of the same individual LiGluK2 receptor diffusing at synapses (Homer1c, red) in the states described in (A). Scale bar, 1 μM. (C and D) Summary of median diffusion coefficient (±IQR), immobile fraction, and MSD versus time curves of synaptic LiGluK2 (C) (n trajectories = 140, in 10 neurons from three independent cultures) and extrasynaptic LiGluK2 (D) (n trajectories = 250, in 10 neurons from three independent cultures) in the closed, desensitized, and recovery closed states. (E) Schematic of SPT experiments as in (A), in the continued presence of VGCC blockers (2-APB, D-APV, ω-conotoxin MVIIC, GYKI 53655, nifedipine, and ryanodine; black) delivered after the initial 488-nm illumination (blue). (F) Example trajectories (yellow) of an individual LiGluK2 receptor diffusing over a portion of dendrite (green) in the indicated states. Homer1c indicates synapses (red). Scale bar, 1 μM. (G and H) Summary of median diffusion coefficient, immobile fraction, and MSD versus time curve of synaptic (G) and extrasynaptic (H) LiGluK2 in different states as indicated in (E) (n trajectories : synaptic = 100; extrasyaptic = 206). Unless otherwise stated, data are presented as mean ± SEM, ∗ p

    Techniques Used: Single-particle Tracking, Diffusion-based Assay

    7) Product Images from "Evidence of functional ryanodine receptors in rat mesenteric collecting lymphatic vessels"

    Article Title: Evidence of functional ryanodine receptors in rat mesenteric collecting lymphatic vessels

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    doi: 10.1152/ajpheart.00564.2018

    The ryanodine receptor (RyR)1/RyR3 inhibitor dantrolene does not inhibit rat mesenteric collecting lymphatic contractions or the increased CF and decreased EDD/Max elicited by substance P (SP). Representative trace of diameter over time of an isolated rat mesenteric collecting lymphatic vessel before and after the addition of 10 −5 M dantrolene, which remained in the bath ( A ). Continuation of the same trace, showing the addition of 10 −8 M SP in the presence of dantrolene ( B ). The boxes beneath the traces in A and B indicate 2-min time periods during which the quantitative data used for comparisons in C – H were collected. Comparisons between baseline (BL), after addition of dantrolene, and after addition of SP in the presence of dantrolene were performed for CF ( C ), EDD/MaxD ( D ), ESD/MaxD ( E ), AMP/MaxD ( F ), EF ( G ), and FPF ( H ). P values are shown for comparisons that were found to be significantly different ( P
    Figure Legend Snippet: The ryanodine receptor (RyR)1/RyR3 inhibitor dantrolene does not inhibit rat mesenteric collecting lymphatic contractions or the increased CF and decreased EDD/Max elicited by substance P (SP). Representative trace of diameter over time of an isolated rat mesenteric collecting lymphatic vessel before and after the addition of 10 −5 M dantrolene, which remained in the bath ( A ). Continuation of the same trace, showing the addition of 10 −8 M SP in the presence of dantrolene ( B ). The boxes beneath the traces in A and B indicate 2-min time periods during which the quantitative data used for comparisons in C – H were collected. Comparisons between baseline (BL), after addition of dantrolene, and after addition of SP in the presence of dantrolene were performed for CF ( C ), EDD/MaxD ( D ), ESD/MaxD ( E ), AMP/MaxD ( F ), EF ( G ), and FPF ( H ). P values are shown for comparisons that were found to be significantly different ( P

    Techniques Used: Isolation

    Blockade of ryanodine receptors (RyRs) with ryanodine leads to a loss of phasic contractions and limited substance P (SP)-induced contraction. Representative trace of diameter over time of an isolated rat collecting mesenteric lymphatic vessel treated with 10 −5 M ryanodine ( A ). The continuation of the trace shows a later time period in the same experiment with ryanodine, when 10 −8 M SP was added. The boxes beneath the trace represent the time periods during which the quantitative data used for comparisons in B – D were collected. The mean diameter ( B ), mean tone ( C ), and maximum tone ( D ) for the time periods before addition of SP, 0–30 s after SP addition, and 31–150 s after addition of SP were compared using a repeated measures ANOVA model followed by Dunnett’s comparison with control (baseline period). P values for the comparisons are shown. N = 6 lymphatics. Each lymphatic studied was obtained from a unique rat. NS, not significant.
    Figure Legend Snippet: Blockade of ryanodine receptors (RyRs) with ryanodine leads to a loss of phasic contractions and limited substance P (SP)-induced contraction. Representative trace of diameter over time of an isolated rat collecting mesenteric lymphatic vessel treated with 10 −5 M ryanodine ( A ). The continuation of the trace shows a later time period in the same experiment with ryanodine, when 10 −8 M SP was added. The boxes beneath the trace represent the time periods during which the quantitative data used for comparisons in B – D were collected. The mean diameter ( B ), mean tone ( C ), and maximum tone ( D ) for the time periods before addition of SP, 0–30 s after SP addition, and 31–150 s after addition of SP were compared using a repeated measures ANOVA model followed by Dunnett’s comparison with control (baseline period). P values for the comparisons are shown. N = 6 lymphatics. Each lymphatic studied was obtained from a unique rat. NS, not significant.

    Techniques Used: Isolation

    Quantitative PCR (qPCR) detection of ryanodine receptors (RyRs) in rat mesenteric collecting lymphatic vessels. A : means show relative expression to GAPDH determined by qPCR and using the 2 -ΔCt method in isolated rat mesenteric collecting lymphatics ( N = 7 rats) and rat brain, skeletal muscle, and cardiac muscle (all N = 2 rats). B : gel of the single amplicon products of expected size from the qPCR reactions. Rat mesenteric collecting lymphatics were harvested (30 lymphatic vessels per rat), and total RNA (50 ng) was reverse transcribed into cDNA and qPCR performed with specific primer/probe sets for Ryr1 (Rn01545085_m1), Ryr2 (Rn01470303_m1), Ryr3 (Rn01486097_m1), and GAPDH (Rn01775763_g1). Reactions with no template were also run as controls (NTC). The gel shown is representative of three separate experiments using mesenteric lymphatic samples obtained from three different rats.
    Figure Legend Snippet: Quantitative PCR (qPCR) detection of ryanodine receptors (RyRs) in rat mesenteric collecting lymphatic vessels. A : means show relative expression to GAPDH determined by qPCR and using the 2 -ΔCt method in isolated rat mesenteric collecting lymphatics ( N = 7 rats) and rat brain, skeletal muscle, and cardiac muscle (all N = 2 rats). B : gel of the single amplicon products of expected size from the qPCR reactions. Rat mesenteric collecting lymphatics were harvested (30 lymphatic vessels per rat), and total RNA (50 ng) was reverse transcribed into cDNA and qPCR performed with specific primer/probe sets for Ryr1 (Rn01545085_m1), Ryr2 (Rn01470303_m1), Ryr3 (Rn01486097_m1), and GAPDH (Rn01775763_g1). Reactions with no template were also run as controls (NTC). The gel shown is representative of three separate experiments using mesenteric lymphatic samples obtained from three different rats.

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Isolation, Amplification

    Determination of ryanodine receptor (RyR)2 and RyR3 localization in isolated rat mesenteric collecting lymphatic vessels with immunofluorescence labeling and laser confocal microscopy. A . maximum intensity z-projection (37 confocal slices) of a lymphatic vessel labeled with anti-RyR2 AlexaFluor488-conjugated secondary antibodies and DAPI. The RyR2 labeling appeared mostly in hoop-like patterns in the vessel wall, with some labeling in the adventitial layer (1 example denoted by the small white arrow). B : maximum intensity z-projection (85 confocal slices) of a lymphatic immunolabeled for RyR3 and nuclei. A similar hoop-like pattern was observed with RyR3 immunolabeling, with some adventitial layer labeling (small white arrow). No signal was detected in labeling controls with no primary antibody (data not shown). C and E : labeling of RyR2, smooth muscle actin (SMA), nuclei, and a merge of the three channels in a single confocal slice featuring a cross-section of the collecting lymphatic wall. The orientation is with the adventitial layer on the left and the endothelial layer on the right. RyR2 labeling was predominantly found in the smooth muscle layer, as evidenced by partial colocalization with SMA and RyR2 labeling surrounding the nuclei of smooth muscle cells. D and F : labeling of RyR3, SMA, nuclei, and a merge of the three channels in a single confocal slice featuring a cross-section of the lymphatic wall, oriented with the adventitial layer on the left and endothelium on the right. RyR3 was predominantly found in smooth muscle cells, as evidenced by the partial colocalization with SMA. The small arrows in E and F point toward nuclei of endothelial cells (ECs). A and B : representative of N = 8 labeling experiments each. C – F : representative of N = 4 experiments each. The scale bars in C – F represent 5 µm.
    Figure Legend Snippet: Determination of ryanodine receptor (RyR)2 and RyR3 localization in isolated rat mesenteric collecting lymphatic vessels with immunofluorescence labeling and laser confocal microscopy. A . maximum intensity z-projection (37 confocal slices) of a lymphatic vessel labeled with anti-RyR2 AlexaFluor488-conjugated secondary antibodies and DAPI. The RyR2 labeling appeared mostly in hoop-like patterns in the vessel wall, with some labeling in the adventitial layer (1 example denoted by the small white arrow). B : maximum intensity z-projection (85 confocal slices) of a lymphatic immunolabeled for RyR3 and nuclei. A similar hoop-like pattern was observed with RyR3 immunolabeling, with some adventitial layer labeling (small white arrow). No signal was detected in labeling controls with no primary antibody (data not shown). C and E : labeling of RyR2, smooth muscle actin (SMA), nuclei, and a merge of the three channels in a single confocal slice featuring a cross-section of the collecting lymphatic wall. The orientation is with the adventitial layer on the left and the endothelial layer on the right. RyR2 labeling was predominantly found in the smooth muscle layer, as evidenced by partial colocalization with SMA and RyR2 labeling surrounding the nuclei of smooth muscle cells. D and F : labeling of RyR3, SMA, nuclei, and a merge of the three channels in a single confocal slice featuring a cross-section of the lymphatic wall, oriented with the adventitial layer on the left and endothelium on the right. RyR3 was predominantly found in smooth muscle cells, as evidenced by the partial colocalization with SMA. The small arrows in E and F point toward nuclei of endothelial cells (ECs). A and B : representative of N = 8 labeling experiments each. C – F : representative of N = 4 experiments each. The scale bars in C – F represent 5 µm.

    Techniques Used: Isolation, Immunofluorescence, Labeling, Confocal Microscopy, Immunolabeling

    Impact of 10 −5 M ryanodine on [Ca 2+ ] i in isolated rat mesenteric collecting lymphatic vessels. A : traces of the F340/F380 ratio obtained over time from a single rat mesenteric collecting lymphatic vessel loaded with the Ca 2+ indicator dye fura-2 AM before and after exposure to 10 −5 M ryanodine (added at time 0 min). Changes in F340/F380 are indicative of changes in [Ca 2+ ] i . B : measurements of luminal diameter obtained from the same lymphatic vessel during the same time periods. C : mean basal F340/F380 between Ca 2+ transients from four collecting lymphatics during the baseline (BL) period before ryanodine treatment and the indicated times after ryanodine was added. Basal F340/F380 significantly increased over BL at 10–11 and 15–16 min after addition of ryanodine. D : mean frequency of Ca 2+ transients, showing a significant mean reduction at 10–11 and 15–16 min after ryanodine was added. E : mean amplitude of the changes in F340/F380 for Ca 2+ transients was significantly decreased at 15–16 min after ryanodine was added, mainly because of the disappearance of Ca 2+ transients at this time point. F : basal diameter/MaxD (EDD/MaxD when phasic contractions were present) significantly decreased form BL at all time points after ryanodine was added. G : amplitude of phasic contractions significantly decreased from BL starting at 6–7 min after ryanodine was added. Lymphatics displaying no phasic contractions were assigned an AMP/MaxD value of 0 in this analysis. Data were analyzed by one-way repeated measures ANOVA followed by Dunnett’s test for multiple comparisons with control. P values for all significant difference are shown. N = 4 isolated mesenteric collecting lymphatics obtained from four different rats. EDD, end-diastolic diameter; MaxD, maximal passive diameter; NS, not significant.
    Figure Legend Snippet: Impact of 10 −5 M ryanodine on [Ca 2+ ] i in isolated rat mesenteric collecting lymphatic vessels. A : traces of the F340/F380 ratio obtained over time from a single rat mesenteric collecting lymphatic vessel loaded with the Ca 2+ indicator dye fura-2 AM before and after exposure to 10 −5 M ryanodine (added at time 0 min). Changes in F340/F380 are indicative of changes in [Ca 2+ ] i . B : measurements of luminal diameter obtained from the same lymphatic vessel during the same time periods. C : mean basal F340/F380 between Ca 2+ transients from four collecting lymphatics during the baseline (BL) period before ryanodine treatment and the indicated times after ryanodine was added. Basal F340/F380 significantly increased over BL at 10–11 and 15–16 min after addition of ryanodine. D : mean frequency of Ca 2+ transients, showing a significant mean reduction at 10–11 and 15–16 min after ryanodine was added. E : mean amplitude of the changes in F340/F380 for Ca 2+ transients was significantly decreased at 15–16 min after ryanodine was added, mainly because of the disappearance of Ca 2+ transients at this time point. F : basal diameter/MaxD (EDD/MaxD when phasic contractions were present) significantly decreased form BL at all time points after ryanodine was added. G : amplitude of phasic contractions significantly decreased from BL starting at 6–7 min after ryanodine was added. Lymphatics displaying no phasic contractions were assigned an AMP/MaxD value of 0 in this analysis. Data were analyzed by one-way repeated measures ANOVA followed by Dunnett’s test for multiple comparisons with control. P values for all significant difference are shown. N = 4 isolated mesenteric collecting lymphatics obtained from four different rats. EDD, end-diastolic diameter; MaxD, maximal passive diameter; NS, not significant.

    Techniques Used: Isolation

    8) Product Images from "Contributions of cardiac “funny” (f) channels and sarcoplasmic reticulum Ca2+ in regulating beating rate of mouse and guinea pig sinoatrial nodeContributions of cardiac “funny” (f) channels and sarcoplasmic reticulum Ca2+ in regulating beating rate of mouse and guinea pig sinoatrial node"

    Article Title: Contributions of cardiac “funny” (f) channels and sarcoplasmic reticulum Ca2+ in regulating beating rate of mouse and guinea pig sinoatrial nodeContributions of cardiac “funny” (f) channels and sarcoplasmic reticulum Ca2+ in regulating beating rate of mouse and guinea pig sinoatrial node

    Journal: Physiological Reports

    doi: 10.14814/phy2.12561

    (A) The effect of I(f) inhibition on the spontaneous beating rate of mouse atrial preparations by 1 μ mol/L ZD 7288 ( n = 9), ryanodine receptor inhibition by 30 μ mol/L ryanodine ( n = 6), and by the combined application of 30 μ mol/L ryanodine + 1 μ mol/L ZD 7288 ( n = 6, Total represents cumulative change from baseline, while Change represents a relative change after ryanodine), SERCA inhibition by 100 μ mol/L CPA ( n = 7) and combined application of 100 μ mol/L CPA + 1 μ mol/L ZD 7288 ( n = 6, Total represents cumulative change from baseline, while Change represents a relative change after CPA ). (B) Data presented as percentage change relative to baseline (except for 30 μ mol/L ryanodine + 1 μ mol/L ZD 7288, and 100 μ mol/L CPA + 1 μ mol/L ZD 7288 combinations as the % changes are calculated relative to reduced baseline by ryanodine and CPA , respectively). Data are expressed as mean ± SEM , n = number of experiments. GraphPad prism (version 5.0) software was used to perform statistical analysis including Student's t ‐test (significance level, P
    Figure Legend Snippet: (A) The effect of I(f) inhibition on the spontaneous beating rate of mouse atrial preparations by 1 μ mol/L ZD 7288 ( n = 9), ryanodine receptor inhibition by 30 μ mol/L ryanodine ( n = 6), and by the combined application of 30 μ mol/L ryanodine + 1 μ mol/L ZD 7288 ( n = 6, Total represents cumulative change from baseline, while Change represents a relative change after ryanodine), SERCA inhibition by 100 μ mol/L CPA ( n = 7) and combined application of 100 μ mol/L CPA + 1 μ mol/L ZD 7288 ( n = 6, Total represents cumulative change from baseline, while Change represents a relative change after CPA ). (B) Data presented as percentage change relative to baseline (except for 30 μ mol/L ryanodine + 1 μ mol/L ZD 7288, and 100 μ mol/L CPA + 1 μ mol/L ZD 7288 combinations as the % changes are calculated relative to reduced baseline by ryanodine and CPA , respectively). Data are expressed as mean ± SEM , n = number of experiments. GraphPad prism (version 5.0) software was used to perform statistical analysis including Student's t ‐test (significance level, P

    Techniques Used: Inhibition, Software

    9) Product Images from "Cerebellar Kainate Receptor-Mediated Facilitation of Glutamate Release Requires Ca2+-Calmodulin and PKA"

    Article Title: Cerebellar Kainate Receptor-Mediated Facilitation of Glutamate Release Requires Ca2+-Calmodulin and PKA

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2018.00195

    Facilitation of glutamate release mediated by presynaptic kainate receptor (KAR) activation requires an increase of Ca 2+ in the cytosol at PF-PuC synapses. (A) Time-course of KA (3 μM) effect on eEPSCs amplitude in control condition (circles) and in slices treated with philanthotoxin (squares). (B) Quantification of modulation observed in (A) . (C) Time-course of the effect of KA on eEPSCs amplitude in control slices (circles) and in thapsigargin-treated slices (squares). (D) In slices treated with thapsigargin or ryanodine, the increase of eEPSCs amplitude induced by KA is prevented. The number of slices (from two to three mice) is indicated in parenthesis at the top of each bar. Results are expressed as means ± SEM (** P
    Figure Legend Snippet: Facilitation of glutamate release mediated by presynaptic kainate receptor (KAR) activation requires an increase of Ca 2+ in the cytosol at PF-PuC synapses. (A) Time-course of KA (3 μM) effect on eEPSCs amplitude in control condition (circles) and in slices treated with philanthotoxin (squares). (B) Quantification of modulation observed in (A) . (C) Time-course of the effect of KA on eEPSCs amplitude in control slices (circles) and in thapsigargin-treated slices (squares). (D) In slices treated with thapsigargin or ryanodine, the increase of eEPSCs amplitude induced by KA is prevented. The number of slices (from two to three mice) is indicated in parenthesis at the top of each bar. Results are expressed as means ± SEM (** P

    Techniques Used: Activation Assay, Mouse Assay

    10) Product Images from "Interactive HIV-1 Tat and Morphine-Induced Synaptodendritic Injury Is Triggered through Focal Disruptions in Na+ Influx, Mitochondrial Instability, and Ca2+ Overload"

    Article Title: Interactive HIV-1 Tat and Morphine-Induced Synaptodendritic Injury Is Triggered through Focal Disruptions in Na+ Influx, Mitochondrial Instability, and Ca2+ Overload

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.5351-13.2014

    Effects of morphine and/or Tat on [Ca 2+ ] i and [Na + ] i in neuronal dendrites following 0–30 min exposure. A , Acute Tat + morphine-induced increases in [Ca 2+ ] i are attenuated by ryanodine or pyruvate, while nimodipine (L-type Ca 2+ channel blocker)
    Figure Legend Snippet: Effects of morphine and/or Tat on [Ca 2+ ] i and [Na + ] i in neuronal dendrites following 0–30 min exposure. A , Acute Tat + morphine-induced increases in [Ca 2+ ] i are attenuated by ryanodine or pyruvate, while nimodipine (L-type Ca 2+ channel blocker)

    Techniques Used:

    11) Product Images from "Enhancement of dendritic persistent Na+ currents by mGluR5 leads to an advancement of spike timing with an increase in temporal precision"

    Article Title: Enhancement of dendritic persistent Na+ currents by mGluR5 leads to an advancement of spike timing with an increase in temporal precision

    Journal: Molecular Brain

    doi: 10.1186/s13041-018-0410-7

    DHPG-induced increase of intrinsic excitability requires intracellular Ca 2+ . a Left, representative traces demonstrate the effects of DHPG on AP firing frequency and afterpotentials at 120 pA current injections when ryanodine (20 μM) was added to the pipette solutions. Right, FI curve summarizes the effects of DHPG on AP firing frequency in response to square pulse injections ( n = 4). b – d Summary plots of the FI curve under the conditions indicated ( b , n = 4, 5 mM nicotinamide; c , n = 5, 100 μM 8-NH 2 -cADPR; d , n = 5, 3 μM calmidazolium). e Bar graphs summarize the effects of DHPG on afterpotentials following 160 pA square current injections under different conditions indicated (n.s., P > 0.05, * P
    Figure Legend Snippet: DHPG-induced increase of intrinsic excitability requires intracellular Ca 2+ . a Left, representative traces demonstrate the effects of DHPG on AP firing frequency and afterpotentials at 120 pA current injections when ryanodine (20 μM) was added to the pipette solutions. Right, FI curve summarizes the effects of DHPG on AP firing frequency in response to square pulse injections ( n = 4). b – d Summary plots of the FI curve under the conditions indicated ( b , n = 4, 5 mM nicotinamide; c , n = 5, 100 μM 8-NH 2 -cADPR; d , n = 5, 3 μM calmidazolium). e Bar graphs summarize the effects of DHPG on afterpotentials following 160 pA square current injections under different conditions indicated (n.s., P > 0.05, * P

    Techniques Used: Transferring

    12) Product Images from "Endoplasmic reticulum stress alters ryanodine receptor function in the murine pancreatic β cell"

    Article Title: Endoplasmic reticulum stress alters ryanodine receptor function in the murine pancreatic β cell

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.RA118.005683

    Ca 2+ signaling and cell death were rescued by ryanodine treatment in islets from Akita mice A and B , glucose-stimulated calcium oscillations were measured in islets isolated from Akita mice treated with DMSO or Ry for 24 h and WT littermate mice treated with DMSO for 24 h. Shown are representative recordings from four individual islets for Akita ( A ) and WT ( B ) mice. C and D , the frequency of oscillations ( C ) and baseline corrected area under curve for calcium responses ( D ) were quantified from three biological replicates per conditions. E , representative pictures of live ( green ) and dead ( red ) staining performed in Akita islets treated with DMSO or Ry for 48 h. Scale bar = 100 μm. F , quantification of the % of dead cells from three repeated experiments. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001 for comparisons between indicated groups. G and H , overall model. G , our data indicate that under ER stress conditions, RyR function is disrupted, leading to increased ER Ca 2+ leak, decreased ER Ca 2+ storage, and altered ER Ca 2+ dynamics. As a consequence, cellular excitability and GICOs are disrupted and activation of the UPR is increased, eventually leading to cell death. H , inhibition of RyR-mediated loss of ER Ca 2 leads to a partial rescue of ER Ca 2+ dynamics under ER stress conditions, which improved cellular excitability and GICOs, delayed initiation of the UPR, and decreased β-cell death. Error bars indicate ± S.D.
    Figure Legend Snippet: Ca 2+ signaling and cell death were rescued by ryanodine treatment in islets from Akita mice A and B , glucose-stimulated calcium oscillations were measured in islets isolated from Akita mice treated with DMSO or Ry for 24 h and WT littermate mice treated with DMSO for 24 h. Shown are representative recordings from four individual islets for Akita ( A ) and WT ( B ) mice. C and D , the frequency of oscillations ( C ) and baseline corrected area under curve for calcium responses ( D ) were quantified from three biological replicates per conditions. E , representative pictures of live ( green ) and dead ( red ) staining performed in Akita islets treated with DMSO or Ry for 48 h. Scale bar = 100 μm. F , quantification of the % of dead cells from three repeated experiments. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001 for comparisons between indicated groups. G and H , overall model. G , our data indicate that under ER stress conditions, RyR function is disrupted, leading to increased ER Ca 2+ leak, decreased ER Ca 2+ storage, and altered ER Ca 2+ dynamics. As a consequence, cellular excitability and GICOs are disrupted and activation of the UPR is increased, eventually leading to cell death. H , inhibition of RyR-mediated loss of ER Ca 2 leads to a partial rescue of ER Ca 2+ dynamics under ER stress conditions, which improved cellular excitability and GICOs, delayed initiation of the UPR, and decreased β-cell death. Error bars indicate ± S.D.

    Techniques Used: Mouse Assay, Isolation, Staining, Activation Assay, Inhibition

    Ryanodine treatment restored calcium oscillations in mouse islets. Glucose-stimulated calcium oscillations were measured in islets isolated from C57BL/6J mice treated with DMSO (control), TM, or TM with Ry for 48 h. A , representative recordings from five single WT islets per condition are shown. B and C , baseline corrected area under the curve was calculated from ( B ) 5 to 30 min (G5) and from ( C ) 30 to 60 min (G15). n = 16–25 islets total from two biological replicates in one experiment; *, p ≤ 0.05; ***, p ≤ 0.001 for comparisons between indicated groups. Error bars indicate ± S.D.
    Figure Legend Snippet: Ryanodine treatment restored calcium oscillations in mouse islets. Glucose-stimulated calcium oscillations were measured in islets isolated from C57BL/6J mice treated with DMSO (control), TM, or TM with Ry for 48 h. A , representative recordings from five single WT islets per condition are shown. B and C , baseline corrected area under the curve was calculated from ( B ) 5 to 30 min (G5) and from ( C ) 30 to 60 min (G15). n = 16–25 islets total from two biological replicates in one experiment; *, p ≤ 0.05; ***, p ≤ 0.001 for comparisons between indicated groups. Error bars indicate ± S.D.

    Techniques Used: Isolation, Mouse Assay

    13) Product Images from "Neuronal Expression of the Human Neuropeptide S Receptor NPSR1 Identifies NPS-Induced Calcium Signaling Pathways"

    Article Title: Neuronal Expression of the Human Neuropeptide S Receptor NPSR1 Identifies NPS-Induced Calcium Signaling Pathways

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0117319

    Model for the intracellular mechanisms underlying NPSR1 activation in cultured mouse hippocampus neurons. Calcium is released from the endoplasmic reticulum via IP 3 and ryanodine receptors, which can be blocked by U73122, 2-APB and ryanodine, respectively. This decrease in the ER calcium content activates store-operated calcium entry (SOCE), which can be visualized by using Ca 2+ -free extracellular solution, the underlying signal cascade can be blocked by ML-9 and SKF96365.
    Figure Legend Snippet: Model for the intracellular mechanisms underlying NPSR1 activation in cultured mouse hippocampus neurons. Calcium is released from the endoplasmic reticulum via IP 3 and ryanodine receptors, which can be blocked by U73122, 2-APB and ryanodine, respectively. This decrease in the ER calcium content activates store-operated calcium entry (SOCE), which can be visualized by using Ca 2+ -free extracellular solution, the underlying signal cascade can be blocked by ML-9 and SKF96365.

    Techniques Used: Activation Assay, Cell Culture

    NPSR1-mediated IP 3 receptor activation triggers calcium-induced calcium release via ryanodine receptors. (A) Fluorescence recording from a single neuron treated with ryanodine [50 μM] (black) and an untreated control (grey). (B) Mean fluorescence intensities calculated from 136 individual neurons. (C) Mean amplitudes calculated from the dataset shown in (B). (D) Fluorescence as averaged from 155 control neurons for two consecutive NPS applications in the absence of ryanodine. (E) Mean amplitudes calculated from the dataset shown in (D). (F) Mean amplitudes for the second application of NPS normalized to the preceding first NPS-administration in control cells (grey) and in the presence of ryanodine (black). NPS was used at 500 nM in all experiments.
    Figure Legend Snippet: NPSR1-mediated IP 3 receptor activation triggers calcium-induced calcium release via ryanodine receptors. (A) Fluorescence recording from a single neuron treated with ryanodine [50 μM] (black) and an untreated control (grey). (B) Mean fluorescence intensities calculated from 136 individual neurons. (C) Mean amplitudes calculated from the dataset shown in (B). (D) Fluorescence as averaged from 155 control neurons for two consecutive NPS applications in the absence of ryanodine. (E) Mean amplitudes calculated from the dataset shown in (D). (F) Mean amplitudes for the second application of NPS normalized to the preceding first NPS-administration in control cells (grey) and in the presence of ryanodine (black). NPS was used at 500 nM in all experiments.

    Techniques Used: Activation Assay, Fluorescence

    14) Product Images from "DA-6034 Induces [Ca2+]i Increase in Epithelial Cells"

    Article Title: DA-6034 Induces [Ca2+]i Increase in Epithelial Cells

    Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

    doi: 10.4196/kjpp.2014.18.2.89

    Relationship with internal Ca 2+ stores and DA-6034. (A) DA-6034 induced [Ca 2+ ] i increases were disappeared after ER depletion by CPA. (B) However, DA-6034 induced Ca 2+ influx not inhibited by the PLC inhibitor, U73122, and its close analogue, U73343. (C) Caffeine induced [Ca 2+ ] i increases were repeated by the second application of caffeine ( upper left panel ) and were blocked by ryanodine, which was an inhibitor of ryanodine receptors ( upper right panel ). DA-6034 induced [Ca 2+ ] i increases also inhibited by ryanodine ( bottom left panel ) and RR ( bottom right panel ). (D) DA-6034 induced [Ca 2+ ] i increases were not inhibited by Baf-A1, which was a specific inhibitor of vacuolar-type H + -ATPase. Data were expressed as the mean±SEM.
    Figure Legend Snippet: Relationship with internal Ca 2+ stores and DA-6034. (A) DA-6034 induced [Ca 2+ ] i increases were disappeared after ER depletion by CPA. (B) However, DA-6034 induced Ca 2+ influx not inhibited by the PLC inhibitor, U73122, and its close analogue, U73343. (C) Caffeine induced [Ca 2+ ] i increases were repeated by the second application of caffeine ( upper left panel ) and were blocked by ryanodine, which was an inhibitor of ryanodine receptors ( upper right panel ). DA-6034 induced [Ca 2+ ] i increases also inhibited by ryanodine ( bottom left panel ) and RR ( bottom right panel ). (D) DA-6034 induced [Ca 2+ ] i increases were not inhibited by Baf-A1, which was a specific inhibitor of vacuolar-type H + -ATPase. Data were expressed as the mean±SEM.

    Techniques Used: Planar Chromatography

    15) Product Images from "Dual action of L-Lactate on the activity of NR2B-containing NMDA receptors: from potentiation to neuroprotection"

    Article Title: Dual action of L-Lactate on the activity of NR2B-containing NMDA receptors: from potentiation to neuroprotection

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-31534-y

    The RyRs blockade impacts only the maintenance of Ca 2+ signal during L-Lactate-induced potentiation. ( A 1 , B 1 ) Averaged Ca 2+ traces recorded from 65 neurons ( A 1 ) and 49 neurons ( B 1 ) following 2 successive applications of glutamate/glycine (2 min; dots) in control and in presence of L-Lactate (10 mM), the neurons in B 1 coming from a culture pre-treated with ryanodine (100 μM). ( A 2 , B 2 ) The scaling of evoked Ca 2+ responses obtained in ( A 1 and B 1 ) shows that the L-Lactate-induced potentiation also corresponds to an increase of the decay time (in comparison with the scaled Ca 2+ response obtained in control, A 2 ), with ryanodine blocking this effect on the Ca 2+ response kinetics ( B 2 ). ( C 1 , C 2 ) Bar charts summarizing the effect of L-Lactate on the Ca 2+ signal triggered by co-application of glutamate/glycine on untreated cultures (n cult = 11; n cells = 468; C 1 ) or cultures pre-treated with ryanodine (n cult = 7; n cells = 331; C 2 ). Note that, when cultures are pre-treated with ryanodine, the potentiating effect of L-Lactate significantly persists only for the amplitude of the Ca 2+ signal. Results are presented as means ± SEM (*p
    Figure Legend Snippet: The RyRs blockade impacts only the maintenance of Ca 2+ signal during L-Lactate-induced potentiation. ( A 1 , B 1 ) Averaged Ca 2+ traces recorded from 65 neurons ( A 1 ) and 49 neurons ( B 1 ) following 2 successive applications of glutamate/glycine (2 min; dots) in control and in presence of L-Lactate (10 mM), the neurons in B 1 coming from a culture pre-treated with ryanodine (100 μM). ( A 2 , B 2 ) The scaling of evoked Ca 2+ responses obtained in ( A 1 and B 1 ) shows that the L-Lactate-induced potentiation also corresponds to an increase of the decay time (in comparison with the scaled Ca 2+ response obtained in control, A 2 ), with ryanodine blocking this effect on the Ca 2+ response kinetics ( B 2 ). ( C 1 , C 2 ) Bar charts summarizing the effect of L-Lactate on the Ca 2+ signal triggered by co-application of glutamate/glycine on untreated cultures (n cult = 11; n cells = 468; C 1 ) or cultures pre-treated with ryanodine (n cult = 7; n cells = 331; C 2 ). Note that, when cultures are pre-treated with ryanodine, the potentiating effect of L-Lactate significantly persists only for the amplitude of the Ca 2+ signal. Results are presented as means ± SEM (*p

    Techniques Used: Blocking Assay

    16) Product Images from "Spontaneous calcium transients manifest in the regenerating muscle and are necessary for skeletal muscle replenishment"

    Article Title: Spontaneous calcium transients manifest in the regenerating muscle and are necessary for skeletal muscle replenishment

    Journal: Cell calcium

    doi: 10.1016/j.ceca.2014.04.004

    Ca 2+ transients are necessary for muscle satellite cell activation and muscle cell precursor proliferation in the regenerating tail. Amputated tadpoles were fixed, coronally sectioned and processed for immunostaining after 24 and 48 hpa with anti-Pax7, muscle satellite cell marker, anti-MyoD, muscle cell precursor marker and anti-PCNA, proliferative marker. (A) Mean ± SEM immunopositive cells per 440 μm thick regenerating tail in control and ryanodine-treated tadpoles at 24 (left) and 48 (right) hpa. *p
    Figure Legend Snippet: Ca 2+ transients are necessary for muscle satellite cell activation and muscle cell precursor proliferation in the regenerating tail. Amputated tadpoles were fixed, coronally sectioned and processed for immunostaining after 24 and 48 hpa with anti-Pax7, muscle satellite cell marker, anti-MyoD, muscle cell precursor marker and anti-PCNA, proliferative marker. (A) Mean ± SEM immunopositive cells per 440 μm thick regenerating tail in control and ryanodine-treated tadpoles at 24 (left) and 48 (right) hpa. *p

    Techniques Used: Activation Assay, Immunostaining, Marker

    Regenerating muscle cells exhibit Ca 2+ transients during the first hours post amputation. Regenerating tail from amputated tadpole was dissociated and cells were cultured, loaded with Fluo4-AM and Ca 2+ -imaged for 1 h at 0.2 Hz acquisition rate followed by immunostaining against the muscle marker 12101. (A) Image shows a representative Ca 2+ -imaged and then immunostained region of field of view. Cells showing Ca 2+ transients during 1 h recording are outlined in red. Green indicates 12101-immunopositive cells. (B) Data show mean ± SEM percent of active muscle cells during the first 10 hpa. (C) Ca 2+ -imaged cells for 30 min were perfused with Ca 2+ -free saline, 5–50 μM ryanodine or vehicle only and imaged for another 30 min followed by immunostaining with the muscle marker 12101. Results show mean ± SEM percent of active muscle cells compared to the 30 min before addition of treatment. *p
    Figure Legend Snippet: Regenerating muscle cells exhibit Ca 2+ transients during the first hours post amputation. Regenerating tail from amputated tadpole was dissociated and cells were cultured, loaded with Fluo4-AM and Ca 2+ -imaged for 1 h at 0.2 Hz acquisition rate followed by immunostaining against the muscle marker 12101. (A) Image shows a representative Ca 2+ -imaged and then immunostained region of field of view. Cells showing Ca 2+ transients during 1 h recording are outlined in red. Green indicates 12101-immunopositive cells. (B) Data show mean ± SEM percent of active muscle cells during the first 10 hpa. (C) Ca 2+ -imaged cells for 30 min were perfused with Ca 2+ -free saline, 5–50 μM ryanodine or vehicle only and imaged for another 30 min followed by immunostaining with the muscle marker 12101. Results show mean ± SEM percent of active muscle cells compared to the 30 min before addition of treatment. *p

    Techniques Used: Cell Culture, Immunostaining, Marker

    Blockade of Ca 2+ transients impairs tail regeneration and muscle replenishment. (A–C) Amputated tadpoles were incubated in vehicle or 50 μM ryanodine for 72 hpa, fixed and photomicrographed (A) followed by whole-mount immunostaining against the muscle marker 12101 (B–C). (B) Maximum intensity projections of the whole-mount 12101-immunostained samples (left) were threshold (right) and a region of interest comprising longitudinally, the tip of the tail on one end, the most posterior organized myotome on the other end, and the full width of the dorsoventral axial musculature, was used to measure the labeled area of the regenerated and differentiated muscle. (C) Maximum intensity projections of whole-mount 12101-immunostained control and treated samples. Values are mean ± SEM percent regenerated muscle compared to control. In A–C, amputation site is indicated with arrowheads. *p
    Figure Legend Snippet: Blockade of Ca 2+ transients impairs tail regeneration and muscle replenishment. (A–C) Amputated tadpoles were incubated in vehicle or 50 μM ryanodine for 72 hpa, fixed and photomicrographed (A) followed by whole-mount immunostaining against the muscle marker 12101 (B–C). (B) Maximum intensity projections of the whole-mount 12101-immunostained samples (left) were threshold (right) and a region of interest comprising longitudinally, the tip of the tail on one end, the most posterior organized myotome on the other end, and the full width of the dorsoventral axial musculature, was used to measure the labeled area of the regenerated and differentiated muscle. (C) Maximum intensity projections of whole-mount 12101-immunostained control and treated samples. Values are mean ± SEM percent regenerated muscle compared to control. In A–C, amputation site is indicated with arrowheads. *p

    Techniques Used: Incubation, Immunostaining, Marker, Labeling

    Proliferation of muscle cell precursors is impaired in ryanodine-treated tadpoles. Ryanodine-treated amputated tadpoles were fixed, sectioned and processed for immunostaining after 24 and 48 hpa. Shown is a representative example of a triple-immunolabeled coronal section of the tail region with anti-Pax7, muscle satellite cell marker in red, anti-MyoD, muscle cell precursor marker in blue and anti-PCNA, proliferative marker in green. Nuclei are labeled with DAPI, in grayscale. A: anterior, P: posterior. Arrowheads indicate amputation site.
    Figure Legend Snippet: Proliferation of muscle cell precursors is impaired in ryanodine-treated tadpoles. Ryanodine-treated amputated tadpoles were fixed, sectioned and processed for immunostaining after 24 and 48 hpa. Shown is a representative example of a triple-immunolabeled coronal section of the tail region with anti-Pax7, muscle satellite cell marker in red, anti-MyoD, muscle cell precursor marker in blue and anti-PCNA, proliferative marker in green. Nuclei are labeled with DAPI, in grayscale. A: anterior, P: posterior. Arrowheads indicate amputation site.

    Techniques Used: Immunostaining, Immunolabeling, Marker, Labeling

    17) Product Images from "Cell Type Specific Spatial and Functional Coupling Between Mammalian Brain Kv2.1 K+ Channels and Ryanodine Receptors"

    Article Title: Cell Type Specific Spatial and Functional Coupling Between Mammalian Brain Kv2.1 K+ Channels and Ryanodine Receptors

    Journal: The Journal of comparative neurology

    doi: 10.1002/cne.23641

    Stimulation of RyR causes a hyperpolarizing shift in the voltage dependence of activation of Kv2.1. A : Typical current recordings of Kv2.1 alone and upon co-expression with the ryanodine receptor 1 (RyR1), ryanodine receptor 2 (RyR2) and ryanodine receptor
    Figure Legend Snippet: Stimulation of RyR causes a hyperpolarizing shift in the voltage dependence of activation of Kv2.1. A : Typical current recordings of Kv2.1 alone and upon co-expression with the ryanodine receptor 1 (RyR1), ryanodine receptor 2 (RyR2) and ryanodine receptor

    Techniques Used: Activation Assay, Expressing

    18) Product Images from "Synaptic crosstalk conferred by a zone of differentially regulated Ca2+ signaling in the dendritic shaft adjoining a potentiated spine"

    Article Title: Synaptic crosstalk conferred by a zone of differentially regulated Ca2+ signaling in the dendritic shaft adjoining a potentiated spine

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1902461116

    Ca 2+ influx through LTCCs initiates CICR via RyRs and promotes STIM1 interaction with LTCCs. ( A ) Superimposed records of LTCC currents measured before and 60 s after the onset of 15-s bath application of Glu. Depolarizations once every 15 s. ( B ) Time course of inhibition of peak inward LTCC current (500-ms steps from −60 to +10 mV, every 15 s) by Glu (15 s) in control (10 mM Ca 2+ ; n = 5), 10 mM Sr 2+ ( n = 5), or 200 μM ryanodine (pretreated for 1 h to block ryanodine receptors; n = 8). Peak LTCC current densities (pA/pF) were calculated by dividing peak inward Ca 2+ current (pA) by cell capacitance (pF). ( C ) Time course of change in [Ca 2+ ] ER evoked by 15-s bath application of Glu in control (10 mM Ca 2+ ; n = 8), 10 mM Sr 2+ ( n = 5), or pretreated with 200 µM ryanodine ( n ) was used to obtain [Ca 2+ ] ER from the D1ER intramolecular FRET ratio, calculated as acceptor-sensitized emission divided by donor emission. ( D ) Time course of change in FRET between STIM1-YFP and CFP-Ca V 1.2 elicited by Glu uncaging for control (3 mM Ca 2+ ; n = 5), 3 mM Sr 2+ ( n = 5), or ryanodine-pretreated neurons ( n = 6). Data are shown as mean (darker) ± SEM (lighter shading). Throughout, mean ± SEM.
    Figure Legend Snippet: Ca 2+ influx through LTCCs initiates CICR via RyRs and promotes STIM1 interaction with LTCCs. ( A ) Superimposed records of LTCC currents measured before and 60 s after the onset of 15-s bath application of Glu. Depolarizations once every 15 s. ( B ) Time course of inhibition of peak inward LTCC current (500-ms steps from −60 to +10 mV, every 15 s) by Glu (15 s) in control (10 mM Ca 2+ ; n = 5), 10 mM Sr 2+ ( n = 5), or 200 μM ryanodine (pretreated for 1 h to block ryanodine receptors; n = 8). Peak LTCC current densities (pA/pF) were calculated by dividing peak inward Ca 2+ current (pA) by cell capacitance (pF). ( C ) Time course of change in [Ca 2+ ] ER evoked by 15-s bath application of Glu in control (10 mM Ca 2+ ; n = 8), 10 mM Sr 2+ ( n = 5), or pretreated with 200 µM ryanodine ( n ) was used to obtain [Ca 2+ ] ER from the D1ER intramolecular FRET ratio, calculated as acceptor-sensitized emission divided by donor emission. ( D ) Time course of change in FRET between STIM1-YFP and CFP-Ca V 1.2 elicited by Glu uncaging for control (3 mM Ca 2+ ; n = 5), 3 mM Sr 2+ ( n = 5), or ryanodine-pretreated neurons ( n = 6). Data are shown as mean (darker) ± SEM (lighter shading). Throughout, mean ± SEM.

    Techniques Used: Inhibition, Mass Spectrometry, Blocking Assay

    19) Product Images from "Multiscale activity imaging in mammary gland reveals how oxytocin enables lactation"

    Article Title: Multiscale activity imaging in mammary gland reveals how oxytocin enables lactation

    Journal: bioRxiv

    doi: 10.1101/657510

    mRNA levels of ryanodine receptors. ( A ) Ryr1 , Ryr2 and Ryr3 expression in lysates prepared from whole mammary tissue (including luminal, basal and stromal cells) dissected from virgin or lactating animals (n = 4 mice). ( B ) Krt14 , Esr1 and Ryr1 levels in freshly sorted luminal and basal cells (n = 3 mice). Graphs show mean ± SEM; * P
    Figure Legend Snippet: mRNA levels of ryanodine receptors. ( A ) Ryr1 , Ryr2 and Ryr3 expression in lysates prepared from whole mammary tissue (including luminal, basal and stromal cells) dissected from virgin or lactating animals (n = 4 mice). ( B ) Krt14 , Esr1 and Ryr1 levels in freshly sorted luminal and basal cells (n = 3 mice). Graphs show mean ± SEM; * P

    Techniques Used: Expressing, Mouse Assay

    20) Product Images from "Alteration of Ryanodine-receptors in Cultured Rat Aortic Smooth Muscle Cells"

    Article Title: Alteration of Ryanodine-receptors in Cultured Rat Aortic Smooth Muscle Cells

    Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

    doi: 10.4196/kjpp.2011.15.6.431

    Ryanodine-induced Ca 2+ release in permeabilized RASMCs. Representative traces of ryanodine-induced Ca 2+ release in freshly dissociated RASMCs (A) and cultured RASMCs (B). (C) Effects of ryanodine on Ca 2+ release in permeabilized freshly dissociated RASMCs and cultured RASMCs. The data, normalized to the initial 10s period prior to ryanodine application, were obtained from four experiments. The arrow indicates the starting points of ryanodine perfusion. Ryanodine (10µM) induced Ca 2+ release from the SR in freshly dissociated RASMCs (■), but not in cultured RASMCs (□).
    Figure Legend Snippet: Ryanodine-induced Ca 2+ release in permeabilized RASMCs. Representative traces of ryanodine-induced Ca 2+ release in freshly dissociated RASMCs (A) and cultured RASMCs (B). (C) Effects of ryanodine on Ca 2+ release in permeabilized freshly dissociated RASMCs and cultured RASMCs. The data, normalized to the initial 10s period prior to ryanodine application, were obtained from four experiments. The arrow indicates the starting points of ryanodine perfusion. Ryanodine (10µM) induced Ca 2+ release from the SR in freshly dissociated RASMCs (■), but not in cultured RASMCs (□).

    Techniques Used: Cell Culture

    21) Product Images from "Acute Simvastatin Inhibits KATP Channels of Porcine Coronary Artery Myocytes"

    Article Title: Acute Simvastatin Inhibits KATP Channels of Porcine Coronary Artery Myocytes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0066404

    Proposed mechanisms for acute simvastatin-induced closure of K ATP channels of vascular myocytes. Simvastatin (lipophilic) crosses the plasma membrane and reaches the sacroplasmic reticulum (SR) of vascular myocytes. Binding of simvastatin to SR leads to the release of ryanodine (Ryr)-sensitive Ca 2+ into the cytosol. Elevation of Ca 2+ activates CaMK II which leads to the subsequent activation (phosphorylation) of AMPKα. Phosphorylation of AMPKα-Thr 172 causes [glucose] o uptake with the participation of SGLT1 and Na + /K + ATPase. Increase in cytosolic [glucose] leads to an elevation of ATP levels via oxidative phosphorylation. Elevation of [ATP] i serves two purposes: (1) closure of vascular K ATP channels, (2) providing phosphate groups for cellular proteins (e.g. PP2A and AMPK) phosphorylation. Phosphorylation of PP2A occurs downstream of AMPK phosphorylation. PP2A phosphorylation results in PP2A inactivation which “releases” AMPK and thus phosphorylation of AMPKα-Thr 172 resulted. AICAR produces similar effects as simvastatin except the initial step involves LKB1-Ser 428 phosphorylation.
    Figure Legend Snippet: Proposed mechanisms for acute simvastatin-induced closure of K ATP channels of vascular myocytes. Simvastatin (lipophilic) crosses the plasma membrane and reaches the sacroplasmic reticulum (SR) of vascular myocytes. Binding of simvastatin to SR leads to the release of ryanodine (Ryr)-sensitive Ca 2+ into the cytosol. Elevation of Ca 2+ activates CaMK II which leads to the subsequent activation (phosphorylation) of AMPKα. Phosphorylation of AMPKα-Thr 172 causes [glucose] o uptake with the participation of SGLT1 and Na + /K + ATPase. Increase in cytosolic [glucose] leads to an elevation of ATP levels via oxidative phosphorylation. Elevation of [ATP] i serves two purposes: (1) closure of vascular K ATP channels, (2) providing phosphate groups for cellular proteins (e.g. PP2A and AMPK) phosphorylation. Phosphorylation of PP2A occurs downstream of AMPK phosphorylation. PP2A phosphorylation results in PP2A inactivation which “releases” AMPK and thus phosphorylation of AMPKα-Thr 172 resulted. AICAR produces similar effects as simvastatin except the initial step involves LKB1-Ser 428 phosphorylation.

    Techniques Used: Binding Assay, Activation Assay

    Role(s) of [Ca 2+ ] o and [Ca 2+ ] i in mediating the effects of simvastatin on AMPK and PP2A activities. (A) Effects of simvastatin (10 μM), with and without ryanodine (100 μM) pre-treatment, on [Ca 2+ ] i changes (F 1 /F 0 ) of porcine coronary artery myocytes, estimated using Fluo-4/AM with confocal laser scanning microscope. (B) Summary of [Ca 2+ ] i changes in response to simvastatin (10 μM) before and after ryanodine (100 μM) challenges. Results are expressed (Area Under Curve, AUC) as mean ± SEM of 13–15 cells (*** P
    Figure Legend Snippet: Role(s) of [Ca 2+ ] o and [Ca 2+ ] i in mediating the effects of simvastatin on AMPK and PP2A activities. (A) Effects of simvastatin (10 μM), with and without ryanodine (100 μM) pre-treatment, on [Ca 2+ ] i changes (F 1 /F 0 ) of porcine coronary artery myocytes, estimated using Fluo-4/AM with confocal laser scanning microscope. (B) Summary of [Ca 2+ ] i changes in response to simvastatin (10 μM) before and after ryanodine (100 μM) challenges. Results are expressed (Area Under Curve, AUC) as mean ± SEM of 13–15 cells (*** P

    Techniques Used: Laser-Scanning Microscopy

    22) Product Images from "H2O2-mediated modulation of cytosolic signaling and organelle function in rat hippocampus"

    Article Title: H2O2-mediated modulation of cytosolic signaling and organelle function in rat hippocampus

    Journal: Pflugers Archiv

    doi: 10.1007/s00424-009-0672-0

    H 2 O 2 releases Ca 2+ from the ER via ryanodine and IP3 receptors. a The ryanodine receptor antagonist dantrolene markedly decreased the amplitude of the H 2 O 2 -induced Ca 2+ rise, and upon wash-out of dantrolene the Ca 2+ transients recovered. In all displayed traces, H 2 O 2 was consistently applied at a concentration of 200 µM. b The IP3 receptor antagonist 2-APB also reversibly depressed the H 2 O 2 -induced Ca 2+ rise. c Combined inhibition of both ryanodine and IP3 receptors almost abolished the H 2 O 2 -induced Ca 2+ rise in a reversible manner. d The sulfhydryl (SH)-protectant dithiothreitol ( DTT ) severely depressed the H 2 O 2 -induced Ca 2+ release, confirming that H 2 O 2 indeed acts via oxidative modulation of SH groups. e Summarized pharmacological profile of the H 2 O 2 (200 µM)-induced Ca 2+ transients. Each drug targeting the ER, ryanodine, or IP3 receptors clearly reduced the H 2 O 2 -induced Ca 2+ rise. All drugs, except for deferoxamine and FeSO 4 , were applied in Ca 2+ -free solutions. Plotted are the normalized Ca 2+ responses, referred to the control responses evoked by H 2 O 2 in each cell before drug treatment. Asterisks mark statistically significant changes (** P
    Figure Legend Snippet: H 2 O 2 releases Ca 2+ from the ER via ryanodine and IP3 receptors. a The ryanodine receptor antagonist dantrolene markedly decreased the amplitude of the H 2 O 2 -induced Ca 2+ rise, and upon wash-out of dantrolene the Ca 2+ transients recovered. In all displayed traces, H 2 O 2 was consistently applied at a concentration of 200 µM. b The IP3 receptor antagonist 2-APB also reversibly depressed the H 2 O 2 -induced Ca 2+ rise. c Combined inhibition of both ryanodine and IP3 receptors almost abolished the H 2 O 2 -induced Ca 2+ rise in a reversible manner. d The sulfhydryl (SH)-protectant dithiothreitol ( DTT ) severely depressed the H 2 O 2 -induced Ca 2+ release, confirming that H 2 O 2 indeed acts via oxidative modulation of SH groups. e Summarized pharmacological profile of the H 2 O 2 (200 µM)-induced Ca 2+ transients. Each drug targeting the ER, ryanodine, or IP3 receptors clearly reduced the H 2 O 2 -induced Ca 2+ rise. All drugs, except for deferoxamine and FeSO 4 , were applied in Ca 2+ -free solutions. Plotted are the normalized Ca 2+ responses, referred to the control responses evoked by H 2 O 2 in each cell before drug treatment. Asterisks mark statistically significant changes (** P

    Techniques Used: Concentration Assay, Inhibition

    23) Product Images from "Modified Cytoplasmic Ca2+ Sequestration Contributes to Spinal Cord Injury-Induced Augmentation of Nerve-Evoked Contractions in the Rat Tail Artery"

    Article Title: Modified Cytoplasmic Ca2+ Sequestration Contributes to Spinal Cord Injury-Induced Augmentation of Nerve-Evoked Contractions in the Rat Tail Artery

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0111804

    Both ryanodine (Ryan; 10 µM) and cyclopiazonic acid (CPZ; 1 µM) increased the blockade of nerve-evoked contractions produced by nifedipine (Nif; 1 µM) in arteries from sham-operated rats (control arteries). By contrast, only ryanodine increased the blockade of nerve-evoked contractions produced by nifedipine in arteries from spinal cord injured rats (SCI arteries). ( A–C ) Averaged overlaid traces showing contractions to 100 pulses at 1 Hz in control ( left traces ) and SCI arteries ( right traces ) in absence ( A ; black line ) or in the presence of ryanodine ( B ; black line ) or CPZ ( C ; black line ) and following addition of nifedipine ( grey lines ). ( D, E ) The % blockade of contractions produced by nifedipine at the 10 th ( D ) and 100 th pulse ( E ) during the trains of stimuli in control ( n = 6) and SCI ( n = 6) arteries in the absence ( white bars ) or in the presence of ryanodine ( grey bars ) or CPZ ( black bars ). Data are presented as means and SEs and statistical comparisons were made with unpaired t -tests. * P
    Figure Legend Snippet: Both ryanodine (Ryan; 10 µM) and cyclopiazonic acid (CPZ; 1 µM) increased the blockade of nerve-evoked contractions produced by nifedipine (Nif; 1 µM) in arteries from sham-operated rats (control arteries). By contrast, only ryanodine increased the blockade of nerve-evoked contractions produced by nifedipine in arteries from spinal cord injured rats (SCI arteries). ( A–C ) Averaged overlaid traces showing contractions to 100 pulses at 1 Hz in control ( left traces ) and SCI arteries ( right traces ) in absence ( A ; black line ) or in the presence of ryanodine ( B ; black line ) or CPZ ( C ; black line ) and following addition of nifedipine ( grey lines ). ( D, E ) The % blockade of contractions produced by nifedipine at the 10 th ( D ) and 100 th pulse ( E ) during the trains of stimuli in control ( n = 6) and SCI ( n = 6) arteries in the absence ( white bars ) or in the presence of ryanodine ( grey bars ) or CPZ ( black bars ). Data are presented as means and SEs and statistical comparisons were made with unpaired t -tests. * P

    Techniques Used: Produced

    Both ryanodine (Ryan; 10 µM) and cyclopiazonic acid (CPZ; 1 µM) increased the amplitude of nerve-evoked contractions in arteries from sham-operated rats (control arteries) but not in those from spinal cord injured rats (SCI arteries). ( A, D ) Averaged traces showing contractions to 100 pulses at 1 Hz in control ( left traces ) and SCI arteries ( right traces ) before ( black line ) and during ( grey line ) application of ryanodine ( A ) or CPZ ( D ). ( B, C, E, F ) Increases in wall tension measured at the 10 th ( B, E ) and 100 th pulse ( C, F ) during the trains of stimuli in control ( n = 6) and SCI ( n = 6) arteries before ( white bars ) and during ( grey bars ) application of ryanodine ( B, C ) or CPZ ( E, F ). Data are presented as means and SEs and statistical comparisons were made with paired t -tests. ** P
    Figure Legend Snippet: Both ryanodine (Ryan; 10 µM) and cyclopiazonic acid (CPZ; 1 µM) increased the amplitude of nerve-evoked contractions in arteries from sham-operated rats (control arteries) but not in those from spinal cord injured rats (SCI arteries). ( A, D ) Averaged traces showing contractions to 100 pulses at 1 Hz in control ( left traces ) and SCI arteries ( right traces ) before ( black line ) and during ( grey line ) application of ryanodine ( A ) or CPZ ( D ). ( B, C, E, F ) Increases in wall tension measured at the 10 th ( B, E ) and 100 th pulse ( C, F ) during the trains of stimuli in control ( n = 6) and SCI ( n = 6) arteries before ( white bars ) and during ( grey bars ) application of ryanodine ( B, C ) or CPZ ( E, F ). Data are presented as means and SEs and statistical comparisons were made with paired t -tests. ** P

    Techniques Used:

    24) Product Images from "Ryanodine receptors selectively contribute to the formation of taste-evoked calcium signals in mouse taste cells"

    Article Title: Ryanodine receptors selectively contribute to the formation of taste-evoked calcium signals in mouse taste cells

    Journal: The European journal of neuroscience

    doi: 10.1111/j.1460-9568.2010.07463.x

    Ryanodine receptors do not contribute to taste-evoked responses in dual responsive taste cells
    Figure Legend Snippet: Ryanodine receptors do not contribute to taste-evoked responses in dual responsive taste cells

    Techniques Used:

    Ryanodine receptors contribute to bitter responses in a sub-population of type II taste cells
    Figure Legend Snippet: Ryanodine receptors contribute to bitter responses in a sub-population of type II taste cells

    Techniques Used:

    Ryanodine receptors contribute to MPG responses in a sub-population of type II taste cells
    Figure Legend Snippet: Ryanodine receptors contribute to MPG responses in a sub-population of type II taste cells

    Techniques Used:

    Ryanodine receptors are functionally expressed in dual-responsive taste cells
    Figure Legend Snippet: Ryanodine receptors are functionally expressed in dual-responsive taste cells

    Techniques Used:

    Ryanodine receptors are functionally expressed in some mouse taste cells
    Figure Legend Snippet: Ryanodine receptors are functionally expressed in some mouse taste cells

    Techniques Used:

    25) Product Images from "Pulsed infrared radiation excites cultured neonatal spiral and vestibular ganglion neurons by modulating mitochondrial calcium cycling"

    Article Title: Pulsed infrared radiation excites cultured neonatal spiral and vestibular ganglion neurons by modulating mitochondrial calcium cycling

    Journal: Journal of Neurophysiology

    doi: 10.1152/jn.00253.2014

    IR modulates mitochondrial Ca 2+ cycling. Treatment was with 50 μM ryanodine (RYN; A ), cyclopiazonic acid (CPA; B ), CGP-37157 (CGP; C ), and ruthenium red (RR; D ). Each graph is a composite that depicts the response of a representative neuron before
    Figure Legend Snippet: IR modulates mitochondrial Ca 2+ cycling. Treatment was with 50 μM ryanodine (RYN; A ), cyclopiazonic acid (CPA; B ), CGP-37157 (CGP; C ), and ruthenium red (RR; D ). Each graph is a composite that depicts the response of a representative neuron before

    Techniques Used:

    26) Product Images from "Ryanodine Receptor-Mediated Calcium Release Has a Key Role in Hippocampal LTD Induction"

    Article Title: Ryanodine Receptor-Mediated Calcium Release Has a Key Role in Hippocampal LTD Induction

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2018.00403

    LTD-induced phosphorylation of CaMKII and Synapsin I. Induction of LTD for 60 min in control (C) slices caused a significant increase in the phosphorylation levels of Synapsin I (left panels), CaMKII-α (center panels) and CaMKII-β (right panels) relative to the levels displayed by unstimulated slices (naïve). Slices pre-incubated for 1 h with 20 μM ryanodine (Rya) before applying the LTD induction protocol displayed significantly lower increments in the phosphorylation levels of Synapsin I and CaMKII-α, whereas the phosphorylation levels of CaMKII-β were not significantly different from the levels displayed by unstimulated slices. Values represent Mean ± SE ( n = 3). Statistical analysis was performed with one-way ANOVA, followed by Tukey’s post hoc test. * p
    Figure Legend Snippet: LTD-induced phosphorylation of CaMKII and Synapsin I. Induction of LTD for 60 min in control (C) slices caused a significant increase in the phosphorylation levels of Synapsin I (left panels), CaMKII-α (center panels) and CaMKII-β (right panels) relative to the levels displayed by unstimulated slices (naïve). Slices pre-incubated for 1 h with 20 μM ryanodine (Rya) before applying the LTD induction protocol displayed significantly lower increments in the phosphorylation levels of Synapsin I and CaMKII-α, whereas the phosphorylation levels of CaMKII-β were not significantly different from the levels displayed by unstimulated slices. Values represent Mean ± SE ( n = 3). Statistical analysis was performed with one-way ANOVA, followed by Tukey’s post hoc test. * p

    Techniques Used: Incubation

    Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on fEPSP rise times, half-widths and decay constants tau measured in CA3–CA1 hippocampal synapses. Left panels: slices were treated with 1 μM ryanodine. (A) fEPSP rise times vs. stimulus intensity. (B) fEPSP half-widths vs. stimulus intensity; (C) fEPSP decay constants (tau) vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine for 60 min. (D) fEPSP rise times vs. stimulus intensity; (E) fEPSP half-widths vs. stimulus intensity; (F) fEPSP decay constants (tau) vs. stimulus intensity. Right panels: slices were treated for 15 min with 1 mM caffeine. (G) fEPSP rise times vs. stimulus intensity; (H) fEPSP half-widths vs. stimulus intensity; (I) fEPSP decay constants (tau) vs. stimulus intensity. Values represent Mean ± SE; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed Mann-Whitney test (* p
    Figure Legend Snippet: Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on fEPSP rise times, half-widths and decay constants tau measured in CA3–CA1 hippocampal synapses. Left panels: slices were treated with 1 μM ryanodine. (A) fEPSP rise times vs. stimulus intensity. (B) fEPSP half-widths vs. stimulus intensity; (C) fEPSP decay constants (tau) vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine for 60 min. (D) fEPSP rise times vs. stimulus intensity; (E) fEPSP half-widths vs. stimulus intensity; (F) fEPSP decay constants (tau) vs. stimulus intensity. Right panels: slices were treated for 15 min with 1 mM caffeine. (G) fEPSP rise times vs. stimulus intensity; (H) fEPSP half-widths vs. stimulus intensity; (I) fEPSP decay constants (tau) vs. stimulus intensity. Values represent Mean ± SE; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed Mann-Whitney test (* p

    Techniques Used: MANN-WHITNEY

    Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on paired-pulse (PP) responses measured in CA3–CA1 hippocampal synapses. (A) Representative fEPSP traces showing PP responses before and after addition of 1 μM ryanodine (left panels), 20 μM ryanodine (center panels) and 1 mM caffeine (right panels). (B) Effects of 1 μM ryanodine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (D) Effects of 1 mM caffeine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (C) Effects of 20 μM ryanodine applied for 60 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. Values represent Mean ± SE; (13, 4) for ryanodine-treated slices; (12, 3) for caffeine-treated slices and (16, 4) for the control. The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used.
    Figure Legend Snippet: Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on paired-pulse (PP) responses measured in CA3–CA1 hippocampal synapses. (A) Representative fEPSP traces showing PP responses before and after addition of 1 μM ryanodine (left panels), 20 μM ryanodine (center panels) and 1 mM caffeine (right panels). (B) Effects of 1 μM ryanodine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (D) Effects of 1 mM caffeine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (C) Effects of 20 μM ryanodine applied for 60 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. Values represent Mean ± SE; (13, 4) for ryanodine-treated slices; (12, 3) for caffeine-treated slices and (16, 4) for the control. The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used.

    Techniques Used:

    Stimulatory and inhibitory ryanodine concentrations and caffeine modify the long-term depression (LTD) response. (A) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the low frequency stimulation (LFS) protocol to control hippocampal slices (14, 4) or to slices treated with 1 μM ryanodine (14, 3) or 20 μM ryanodine (13, 4). Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min after applying the LFS protocol (trace 2) to control slices, or recorded in slices treated with 1 μM or 20 μM ryanodine are shown on top of the graph. Open symbols: control slices; gray symbols: slices treated with 1 μM ryanodine; black symbols: slices treated with 20 μM ryanodine. (B) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation. (C) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the LFS protocol to control hippocampal slices (15, 3) or to slices treated with 1 mM caffeine (12, 4). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min (trace 2) after applying the LFS protocol to control slices and to slices treated with 1 mM caffeine are shown on top of the graph. Open symbols: control slices; black symbols: slices treated with 1 mM caffeine. (D) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation of control or caffeine-treated slices. Values represent Mean ± SE. Statistical significance of values was assessed by Mann-Whitney test (* p
    Figure Legend Snippet: Stimulatory and inhibitory ryanodine concentrations and caffeine modify the long-term depression (LTD) response. (A) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the low frequency stimulation (LFS) protocol to control hippocampal slices (14, 4) or to slices treated with 1 μM ryanodine (14, 3) or 20 μM ryanodine (13, 4). Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min after applying the LFS protocol (trace 2) to control slices, or recorded in slices treated with 1 μM or 20 μM ryanodine are shown on top of the graph. Open symbols: control slices; gray symbols: slices treated with 1 μM ryanodine; black symbols: slices treated with 20 μM ryanodine. (B) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation. (C) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the LFS protocol to control hippocampal slices (15, 3) or to slices treated with 1 mM caffeine (12, 4). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min (trace 2) after applying the LFS protocol to control slices and to slices treated with 1 mM caffeine are shown on top of the graph. Open symbols: control slices; black symbols: slices treated with 1 mM caffeine. (D) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation of control or caffeine-treated slices. Values represent Mean ± SE. Statistical significance of values was assessed by Mann-Whitney test (* p

    Techniques Used: MANN-WHITNEY

    Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on basal synaptic transmission in the CA3–CA1 hippocampal synapse. Left panels: slices were treated with 1 μM ryanodine; representative field excitatory postsynaptic potential (fEPSP) traces registered before and 15 min after ryanodine addition are illustrated on top of the panels. (A) Fiber volley (FV) amplitude vs. stimulus intensity; (B) amplitude vs. stimulus intensity; (C) fEPSP slopes vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine; representative fEPSP traces registered before and 60 min after ryanodine addition are illustrated on top. (D) FV amplitude vs. stimulus intensity; (E) fEPSP amplitude vs. stimulus intensity; (F) fEPSP slopes vs. stimulus intensity. Right Panels: slices were treated with 1 mM caffeine; representative fEPSP traces registered before and 15 min after caffeine addition are illustrated on top. (G) FV amplitude vs. stimulus intensity; (H) fEPSP amplitude vs. stimulus intensity; (I) fEPSP slopes vs. stimulus intensity. Values represent Mean ± SEM; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed by Mann-Whitney test ( p > 0.05 in all cases).
    Figure Legend Snippet: Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on basal synaptic transmission in the CA3–CA1 hippocampal synapse. Left panels: slices were treated with 1 μM ryanodine; representative field excitatory postsynaptic potential (fEPSP) traces registered before and 15 min after ryanodine addition are illustrated on top of the panels. (A) Fiber volley (FV) amplitude vs. stimulus intensity; (B) amplitude vs. stimulus intensity; (C) fEPSP slopes vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine; representative fEPSP traces registered before and 60 min after ryanodine addition are illustrated on top. (D) FV amplitude vs. stimulus intensity; (E) fEPSP amplitude vs. stimulus intensity; (F) fEPSP slopes vs. stimulus intensity. Right Panels: slices were treated with 1 mM caffeine; representative fEPSP traces registered before and 15 min after caffeine addition are illustrated on top. (G) FV amplitude vs. stimulus intensity; (H) fEPSP amplitude vs. stimulus intensity; (I) fEPSP slopes vs. stimulus intensity. Values represent Mean ± SEM; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed by Mann-Whitney test ( p > 0.05 in all cases).

    Techniques Used: Transmission Assay, MANN-WHITNEY

    27) Product Images from "Presynaptic α4β2 Nicotinic Acetylcholine Receptors Increase Glutamate Release and Serotonin Neuron Excitability in the Dorsal Raphe Nucleus"

    Article Title: Presynaptic α4β2 Nicotinic Acetylcholine Receptors Increase Glutamate Release and Serotonin Neuron Excitability in the Dorsal Raphe Nucleus

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0941-12.2012

    Nicotinic effects depend on VGCCs and intracellular CICR. A , Time–frequency histogram shows the effect of nicotine on the sEPSC frequency in the presence of CdCl 2 (gray bar). B , Time–frequency histogram shows the lack of effect of nicotine on the sEPSC frequency in the presence of a mixture containing the Ca 2+ channel blockers ω-agatoxin-TK, ω-conotoxin-GVIA, and nitrendipine (gray bar). C , Time–frequency histogram shows the lack of effect of nicotine on the sEPSC frequency in the presence of the SERCA blocker thapsigargin (gray bar). D , Bar graph shows the effect of nicotine on the sEPSC frequency in slices pretreated with CdCl 2 , Ca 2+ channel blockers, thapsigargin, CPA, or ryanodine (** p
    Figure Legend Snippet: Nicotinic effects depend on VGCCs and intracellular CICR. A , Time–frequency histogram shows the effect of nicotine on the sEPSC frequency in the presence of CdCl 2 (gray bar). B , Time–frequency histogram shows the lack of effect of nicotine on the sEPSC frequency in the presence of a mixture containing the Ca 2+ channel blockers ω-agatoxin-TK, ω-conotoxin-GVIA, and nitrendipine (gray bar). C , Time–frequency histogram shows the lack of effect of nicotine on the sEPSC frequency in the presence of the SERCA blocker thapsigargin (gray bar). D , Bar graph shows the effect of nicotine on the sEPSC frequency in slices pretreated with CdCl 2 , Ca 2+ channel blockers, thapsigargin, CPA, or ryanodine (** p

    Techniques Used:

    Model summarizing nicotinic effects on glutamate terminals in the DRN. A , In physiological conditions, the cholinergic tone is regulating the excitatory glutamatergic input to the DRN 5-HT neurons through the activation of α4β2 nAChRs, located at glutamate terminals. Released ACh is quickly metabolized by the enzyme acetylcholinesterase. As a result, the Ca 2+ influx into the glutamate terminals and glutamate release will be maintained at low levels. B , When nicotine is present (for example, in smokers), more α4β2 nAChRs will be activated, because nicotine cannot be degraded in the synaptic cleft. This will enhance Ca 2+ entry, followed by depolarization of glutamate terminals and activation of VGCCs. This, in turn, will increase even more intracellular calcium and produce CICR from the endoplasmic reticulum (ER) through the activation of ryanodine receptors (RyR). This last event generates a long-term potentiation of glutamate release.
    Figure Legend Snippet: Model summarizing nicotinic effects on glutamate terminals in the DRN. A , In physiological conditions, the cholinergic tone is regulating the excitatory glutamatergic input to the DRN 5-HT neurons through the activation of α4β2 nAChRs, located at glutamate terminals. Released ACh is quickly metabolized by the enzyme acetylcholinesterase. As a result, the Ca 2+ influx into the glutamate terminals and glutamate release will be maintained at low levels. B , When nicotine is present (for example, in smokers), more α4β2 nAChRs will be activated, because nicotine cannot be degraded in the synaptic cleft. This will enhance Ca 2+ entry, followed by depolarization of glutamate terminals and activation of VGCCs. This, in turn, will increase even more intracellular calcium and produce CICR from the endoplasmic reticulum (ER) through the activation of ryanodine receptors (RyR). This last event generates a long-term potentiation of glutamate release.

    Techniques Used: Activation Assay

    28) Product Images from "Multiple forms of LTP in hippocampal CA3 neurons use a common postsynaptic mechanism"

    Article Title: Multiple forms of LTP in hippocampal CA3 neurons use a common postsynaptic mechanism

    Journal: Nature neuroscience

    doi: 10.1038/10180

    Ca 2+ release from internal stores when fast synaptic transmission is blocked. (a) In the presence of kynurenate (8–10 mM), L-HFS-3 elicited a rise in [Ca 2+ ] i . An individual experiment in which the mGluR1 antagonist AIDA (500 μM) blocked the rise in [Ca 2+ ] i during L-HFS. After washout (30 min), a large rise in [Ca 2+ ] i was evoked in the proximal dendrite by L-HFS. In this case, a sustained depolarization was observed both with and without AIDA. (Waveforms are from single stimulation trains.) (b) Ryanodine (ryn; n = 3), thapsigargin (thaps; n = 2), MCPG ( n = 3), AIDA ( n = 6) or CPCCOEt ( n = 3) blocked the rise in postsynaptic [Ca 2+ ], consistent with the block of Ca 2+ release from internal stores. (c) AIDA ( n = 3) or CPCCOEt ( n = 3) blocked mossy fiber LTP when fast synaptic transmission was blocked (83.1 ± 4.3%, n = 6; data were combined for the two drugs). These results were significantly different from LTP induced without mGluR antagonists present ( p
    Figure Legend Snippet: Ca 2+ release from internal stores when fast synaptic transmission is blocked. (a) In the presence of kynurenate (8–10 mM), L-HFS-3 elicited a rise in [Ca 2+ ] i . An individual experiment in which the mGluR1 antagonist AIDA (500 μM) blocked the rise in [Ca 2+ ] i during L-HFS. After washout (30 min), a large rise in [Ca 2+ ] i was evoked in the proximal dendrite by L-HFS. In this case, a sustained depolarization was observed both with and without AIDA. (Waveforms are from single stimulation trains.) (b) Ryanodine (ryn; n = 3), thapsigargin (thaps; n = 2), MCPG ( n = 3), AIDA ( n = 6) or CPCCOEt ( n = 3) blocked the rise in postsynaptic [Ca 2+ ], consistent with the block of Ca 2+ release from internal stores. (c) AIDA ( n = 3) or CPCCOEt ( n = 3) blocked mossy fiber LTP when fast synaptic transmission was blocked (83.1 ± 4.3%, n = 6; data were combined for the two drugs). These results were significantly different from LTP induced without mGluR antagonists present ( p

    Techniques Used: Transmission Assay, Blocking Assay

    29) Product Images from "Synaptic crosstalk conferred by a zone of differentially regulated Ca2+ signaling in the dendritic shaft adjoining a potentiated spine"

    Article Title: Synaptic crosstalk conferred by a zone of differentially regulated Ca2+ signaling in the dendritic shaft adjoining a potentiated spine

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1902461116

    Ca 2+ influx through LTCCs initiates CICR via RyRs and promotes STIM1 interaction with LTCCs. ( A ) Superimposed records of LTCC currents measured before and 60 s after the onset of 15-s bath application of Glu. Depolarizations once every 15 s. ( B ) Time course of inhibition of peak inward LTCC current (500-ms steps from −60 to +10 mV, every 15 s) by Glu (15 s) in control (10 mM Ca 2+ ; n = 5), 10 mM Sr 2+ ( n = 5), or 200 μM ryanodine (pretreated for 1 h to block ryanodine receptors; n = 8). Peak LTCC current densities (pA/pF) were calculated by dividing peak inward Ca 2+ current (pA) by cell capacitance (pF). ( C ) Time course of change in [Ca 2+ ] ER evoked by 15-s bath application of Glu in control (10 mM Ca 2+ ; n = 8), 10 mM Sr 2+ ( n = 5), or pretreated with 200 µM ryanodine ( n ) was used to obtain [Ca 2+ ] ER from the D1ER intramolecular FRET ratio, calculated as acceptor-sensitized emission divided by donor emission. ( D ) Time course of change in FRET between STIM1-YFP and CFP-Ca V 1.2 elicited by Glu uncaging for control (3 mM Ca 2+ ; n = 5), 3 mM Sr 2+ ( n = 5), or ryanodine-pretreated neurons ( n = 6). Data are shown as mean (darker) ± SEM (lighter shading). Throughout, mean ± SEM.
    Figure Legend Snippet: Ca 2+ influx through LTCCs initiates CICR via RyRs and promotes STIM1 interaction with LTCCs. ( A ) Superimposed records of LTCC currents measured before and 60 s after the onset of 15-s bath application of Glu. Depolarizations once every 15 s. ( B ) Time course of inhibition of peak inward LTCC current (500-ms steps from −60 to +10 mV, every 15 s) by Glu (15 s) in control (10 mM Ca 2+ ; n = 5), 10 mM Sr 2+ ( n = 5), or 200 μM ryanodine (pretreated for 1 h to block ryanodine receptors; n = 8). Peak LTCC current densities (pA/pF) were calculated by dividing peak inward Ca 2+ current (pA) by cell capacitance (pF). ( C ) Time course of change in [Ca 2+ ] ER evoked by 15-s bath application of Glu in control (10 mM Ca 2+ ; n = 8), 10 mM Sr 2+ ( n = 5), or pretreated with 200 µM ryanodine ( n ) was used to obtain [Ca 2+ ] ER from the D1ER intramolecular FRET ratio, calculated as acceptor-sensitized emission divided by donor emission. ( D ) Time course of change in FRET between STIM1-YFP and CFP-Ca V 1.2 elicited by Glu uncaging for control (3 mM Ca 2+ ; n = 5), 3 mM Sr 2+ ( n = 5), or ryanodine-pretreated neurons ( n = 6). Data are shown as mean (darker) ± SEM (lighter shading). Throughout, mean ± SEM.

    Techniques Used: Inhibition, Mass Spectrometry, Blocking Assay

    30) Product Images from "Pregnancy increases Ca2+ sparks/STOCs and reduces uterine arterial myogenic tone"

    Article Title: Pregnancy increases Ca2+ sparks/STOCs and reduces uterine arterial myogenic tone

    Journal: Hypertension (Dallas, Tex. : 1979)

    doi: 10.1161/HYPERTENSIONAHA.118.12484

    Pregnancy increases Ca 2+ sparks in uterine arterial myocytes. Ca 2+ sparks were measured in en-face endothelium-denuded uterine arteries from nonpregnant (UA NP ) and pregnant (UA P ) sheep. A and B. Representative line scan images of Fluo-4AM loaded uterine arteries showing Ca 2+ sparks recorded before and after the sequential application of 30 mmol/L K + (30K) and 30 mmol/L K + + 30 μmol/L ryanodine (30K/Ry). C. Percentage firing of Ca 2+ sparks in arterial myocytes under control conditions and after the application of high K + followed by ryanodine are summarized from a total of 1841 line scans in uterine arteries of 6 nonpregnant (control: 327, 30K: 315, and 30K/Ry: 300) and 6 pregnant (control: 299, 30K: 300, and 30K/Ry: 300) animals. D. Bar graph of Ca 2+ spark frequency (sparks/μm/s) before and after the application of 30K and 30K/Ry. Data are means ± S.E.M. of 6 animals of each group. *P
    Figure Legend Snippet: Pregnancy increases Ca 2+ sparks in uterine arterial myocytes. Ca 2+ sparks were measured in en-face endothelium-denuded uterine arteries from nonpregnant (UA NP ) and pregnant (UA P ) sheep. A and B. Representative line scan images of Fluo-4AM loaded uterine arteries showing Ca 2+ sparks recorded before and after the sequential application of 30 mmol/L K + (30K) and 30 mmol/L K + + 30 μmol/L ryanodine (30K/Ry). C. Percentage firing of Ca 2+ sparks in arterial myocytes under control conditions and after the application of high K + followed by ryanodine are summarized from a total of 1841 line scans in uterine arteries of 6 nonpregnant (control: 327, 30K: 315, and 30K/Ry: 300) and 6 pregnant (control: 299, 30K: 300, and 30K/Ry: 300) animals. D. Bar graph of Ca 2+ spark frequency (sparks/μm/s) before and after the application of 30K and 30K/Ry. Data are means ± S.E.M. of 6 animals of each group. *P

    Techniques Used:

    Spontaneous transient outward currents (STOCs) in uterine arterial smooth muscle cells. A and B. STOCs recorded from uterine arterial smooth muscle cells of nonpregnant (UA NP , A ) and pregnant (UA P , B ) sheep. C. Blockade of STOCs by iberiotoxin (100 nmol/L). D. Blockade of STOCs by ryanodine (30 μmol/L). Similar results were obtained in cells of 5–6 animals. HP: holding potential.
    Figure Legend Snippet: Spontaneous transient outward currents (STOCs) in uterine arterial smooth muscle cells. A and B. STOCs recorded from uterine arterial smooth muscle cells of nonpregnant (UA NP , A ) and pregnant (UA P , B ) sheep. C. Blockade of STOCs by iberiotoxin (100 nmol/L). D. Blockade of STOCs by ryanodine (30 μmol/L). Similar results were obtained in cells of 5–6 animals. HP: holding potential.

    Techniques Used:

    Pregnancy upregulates ryanodine receptors in uterine arteries. Ryanodine receptor type 1 (RyR1), type 2 (RyR2) and type 3 (RyR3) mRNA ( A ) and protein ( B ) abundance in uterine arteries of nonpregnant (UA NP ) and pregnant (UA P ) sheep were determined with real-time RT-PCR and Western blotting. Data are means ± SEM from 4–5 animals of each group. *P
    Figure Legend Snippet: Pregnancy upregulates ryanodine receptors in uterine arteries. Ryanodine receptor type 1 (RyR1), type 2 (RyR2) and type 3 (RyR3) mRNA ( A ) and protein ( B ) abundance in uterine arteries of nonpregnant (UA NP ) and pregnant (UA P ) sheep were determined with real-time RT-PCR and Western blotting. Data are means ± SEM from 4–5 animals of each group. *P

    Techniques Used: Quantitative RT-PCR, Western Blot

    31) Product Images from "Inhibitory effect of Suaeda asparagoides (Miq.) extract on the motility of rat gastric antrum is mediated by ?-adrenoceptor"

    Article Title: Inhibitory effect of Suaeda asparagoides (Miq.) extract on the motility of rat gastric antrum is mediated by ?-adrenoceptor

    Journal: Laboratory Animal Research

    doi: 10.5625/lar.2011.27.4.317

    Effect of ryanodine on SAWF induced inhibition on amplitude of rat gastric antral muscle phasic contraction. (A) Panel for SAWF induced inhibition on the amplitude of spontaneous phasic contraction (B) Panel for the effect of SAWF on ryanodine pretreated antral muscle strip. The bars over the panels indicate the duration of administration of the extract ryanodine. (C) Summary for the control, SAWF and ryanidine treatment groups. (D) Ryanodine (10 µM) did not affect SAWF induced inhibition ratio on the amplitude of rat antral muscle phasic contraction. Values are mean±SEM. * Significantly different from previous column, P
    Figure Legend Snippet: Effect of ryanodine on SAWF induced inhibition on amplitude of rat gastric antral muscle phasic contraction. (A) Panel for SAWF induced inhibition on the amplitude of spontaneous phasic contraction (B) Panel for the effect of SAWF on ryanodine pretreated antral muscle strip. The bars over the panels indicate the duration of administration of the extract ryanodine. (C) Summary for the control, SAWF and ryanidine treatment groups. (D) Ryanodine (10 µM) did not affect SAWF induced inhibition ratio on the amplitude of rat antral muscle phasic contraction. Values are mean±SEM. * Significantly different from previous column, P

    Techniques Used: Inhibition, Stripping Membranes

    32) Product Images from "RGS2 squelches vascular Gi/o and Gq signaling to modulate myogenic tone and promote uterine blood flow. RGS2 squelches vascular Gi/o and Gq signaling to modulate myogenic tone and promote uterine blood flow"

    Article Title: RGS2 squelches vascular Gi/o and Gq signaling to modulate myogenic tone and promote uterine blood flow. RGS2 squelches vascular Gi/o and Gq signaling to modulate myogenic tone and promote uterine blood flow

    Journal: Physiological Reports

    doi: 10.14814/phy2.12692

    Increased Ca 2+ release from internal stores mediates augmented myogenic tone in RGS 2 deficient mice. (A B) Myogenic response of uterine arteries from wild type ( Rgs2+/+ ) and Rgs2 knockout ( Rgs2−/− ) mice in the absence and presence of ryanodine (Ryan, closed triangles), thapsigargin (Thaps, open diamonds), or Ryan plus N,N,N′,N′‐tetrakis(2‐pyridylmethyl)ethane‐1,2‐diamine (Ryan + TPEN , open squares). Thaps and Ryan + TPEN almost completely blocked myogenic response in both genotypes. (C) Depletion of extracellular Ca 2+ with Ca 2+ ‐free PSS induces robust Ca 2+ release from internal stores in uterine artery smooth muscle cells ( SMC s) from Rgs2 knockout ( Rgs2−/− , open circles) relative and wild‐type ( Rgs2+/+ , closed circles) mice. Ionomycin induces Ca 2+ release from internal stores in SMC s from both genotypes. (D) Ca 2+ ‐free PSS but not ionomycin‐induced Ca 2+ release from internal stores is almost completely abolished by Ryan in SMC s from Rgs2−/− mice. (E) A plot of area under the curve ( AUC ) of fura‐2 fluorescence shown in C and D. Note the augmentation of ionomycin‐induced Ca 2+ response by Ryan in Rgs2+/+ SMC s, which correlates with enhanced myogenic response in Rgs2+/+ uterine arteries in the presence of Ryan in A. Values are mean ± SE. * , ** P
    Figure Legend Snippet: Increased Ca 2+ release from internal stores mediates augmented myogenic tone in RGS 2 deficient mice. (A B) Myogenic response of uterine arteries from wild type ( Rgs2+/+ ) and Rgs2 knockout ( Rgs2−/− ) mice in the absence and presence of ryanodine (Ryan, closed triangles), thapsigargin (Thaps, open diamonds), or Ryan plus N,N,N′,N′‐tetrakis(2‐pyridylmethyl)ethane‐1,2‐diamine (Ryan + TPEN , open squares). Thaps and Ryan + TPEN almost completely blocked myogenic response in both genotypes. (C) Depletion of extracellular Ca 2+ with Ca 2+ ‐free PSS induces robust Ca 2+ release from internal stores in uterine artery smooth muscle cells ( SMC s) from Rgs2 knockout ( Rgs2−/− , open circles) relative and wild‐type ( Rgs2+/+ , closed circles) mice. Ionomycin induces Ca 2+ release from internal stores in SMC s from both genotypes. (D) Ca 2+ ‐free PSS but not ionomycin‐induced Ca 2+ release from internal stores is almost completely abolished by Ryan in SMC s from Rgs2−/− mice. (E) A plot of area under the curve ( AUC ) of fura‐2 fluorescence shown in C and D. Note the augmentation of ionomycin‐induced Ca 2+ response by Ryan in Rgs2+/+ SMC s, which correlates with enhanced myogenic response in Rgs2+/+ uterine arteries in the presence of Ryan in A. Values are mean ± SE. * , ** P

    Techniques Used: Mouse Assay, Knock-Out, Fluorescence

    A model summarizing smooth muscle signaling pathways that are regulated by RGS 2 to control uterine artery myogenic tone. Under conditions where RGS 2 is absent or deficient, G protein activation by mechanosensation leads to a sustained phospholipase C ( PLC )‐mediated Ca 2+ release from the sarcoplasmic reticulum ( SR ), which increases cytosolic Ca 2+ concentration and activates Ca 2+ ‐Cam/ MLCK ‐mediated smooth muscle contraction. High cytosolic Ca 2+ due to the absence/deficiency of RGS 2 also inhibits SR Ca 2+ release via ryanodine receptors (RyR), which normally inhibits contraction by promoting membrane hyperpolarization. Cam, calmodulin; MLCK , myosin light‐chain kinase; BK C a 2+ , Ca 2+ ‐activated potassium channel; myosin‐P, phosphorylated myosin.
    Figure Legend Snippet: A model summarizing smooth muscle signaling pathways that are regulated by RGS 2 to control uterine artery myogenic tone. Under conditions where RGS 2 is absent or deficient, G protein activation by mechanosensation leads to a sustained phospholipase C ( PLC )‐mediated Ca 2+ release from the sarcoplasmic reticulum ( SR ), which increases cytosolic Ca 2+ concentration and activates Ca 2+ ‐Cam/ MLCK ‐mediated smooth muscle contraction. High cytosolic Ca 2+ due to the absence/deficiency of RGS 2 also inhibits SR Ca 2+ release via ryanodine receptors (RyR), which normally inhibits contraction by promoting membrane hyperpolarization. Cam, calmodulin; MLCK , myosin light‐chain kinase; BK C a 2+ , Ca 2+ ‐activated potassium channel; myosin‐P, phosphorylated myosin.

    Techniques Used: Activation Assay, Planar Chromatography, Concentration Assay, Chick Chorioallantoic Membrane Assay

    33) Product Images from "Clustering of Ca2+ transients in interstitial cells of Cajal defines slow wave duration"

    Article Title: Clustering of Ca2+ transients in interstitial cells of Cajal defines slow wave duration

    Journal: The Journal of General Physiology

    doi: 10.1085/jgp.201711771

    Mechanism of slow wave propagation via asynchronous Ca 2+ release. Schematic showing the mechanism by which slow waves in the mouse small intestine propagate via asynchronous ER Ca 2+ release. Electrical slow waves are propagating depolarizing events, which are actively propagated through ICC-MY networks. (1) When a slow wave propagates through the ICC-MY network, the resulting depolarization activates voltage-gated T-type Ca 2+ channels on the plasma membrane. (2) Ca 2+ influx from the opening of T-type Ca 2+ channels allows Ca 2+ ions to enter an excluded volume or microdomain and can then activate Ca 2+ release channels such as RyRs (with amplification from IP 3 Rs) on the membrane of the ER, possibly via a CICR mechanism. (3) Multiple sites of RyRs/IP 3 Rs are located across multiple microdomains in a given single ICC-MY, and thus Ca 2+ release occurs from multiple sites. Because of the excluded volume of the microdomain, individual Ca 2+ release events manifests as temporally brief events that also occur asynchronously among each other, leading to a summated Ca 2+ signal from the total sites in a cell lasting ∼1 s. (4) The summated Ca 2+ signal resulting from multiple Ca 2+ release events is able to sustain ANO1 activation, resulting in prolonged Cl − efflux from the cell and a corresponding sustained depolarization for the duration of the slow wave plateau. RyR, ryanodine receptor; PM, plasma membrane; CICR, calcium-induced calcium release.
    Figure Legend Snippet: Mechanism of slow wave propagation via asynchronous Ca 2+ release. Schematic showing the mechanism by which slow waves in the mouse small intestine propagate via asynchronous ER Ca 2+ release. Electrical slow waves are propagating depolarizing events, which are actively propagated through ICC-MY networks. (1) When a slow wave propagates through the ICC-MY network, the resulting depolarization activates voltage-gated T-type Ca 2+ channels on the plasma membrane. (2) Ca 2+ influx from the opening of T-type Ca 2+ channels allows Ca 2+ ions to enter an excluded volume or microdomain and can then activate Ca 2+ release channels such as RyRs (with amplification from IP 3 Rs) on the membrane of the ER, possibly via a CICR mechanism. (3) Multiple sites of RyRs/IP 3 Rs are located across multiple microdomains in a given single ICC-MY, and thus Ca 2+ release occurs from multiple sites. Because of the excluded volume of the microdomain, individual Ca 2+ release events manifests as temporally brief events that also occur asynchronously among each other, leading to a summated Ca 2+ signal from the total sites in a cell lasting ∼1 s. (4) The summated Ca 2+ signal resulting from multiple Ca 2+ release events is able to sustain ANO1 activation, resulting in prolonged Cl − efflux from the cell and a corresponding sustained depolarization for the duration of the slow wave plateau. RyR, ryanodine receptor; PM, plasma membrane; CICR, calcium-induced calcium release.

    Techniques Used: Immunocytochemistry, Amplification, Activation Assay

    The effect of ryanodine on Ca 2+ transients in ICC-MY. (A and B) Representative heat map showing the summated PTCLs for an entire 60× magnification recording of ICC-MY in control and ryanodine (100 µM) showing the inhibitory effects of ryanodine (A), as indicated in the occurrence map of individually color-coded Ca 2+ firing sites in the ICC-MY network (B). (C) Traces of PTCL activity (PTCL area [dark blue] and PTCL count [brown]) in control and ryanodine. (D and E) Histogram showing the probability (%) that an individual Ca 2+ firing site in the ICC-MY cell somata and cell processes in E will fire during a CTC cycle in control conditions (black bars) and ryanodine (100 µM; red bars; n = 5, FOV = 5). (F) The number of Ca 2+ firing sites per cell soma was reduced from 6.28 ± 1.47 in control to 0.94 ± 0.81 in the presence of ryanodine (100 µM; P = 0.01, n = 5, FOV = 5). (G) PTCL area/frame was reduced in the cell somata from 62.19 ± 6.34 µm 2 in control to 0.3 ± 0.2 µm 2 in ryanodine (100 µM; P = 0.002, n = 5, FOV = 5). (H) PTCL count/frame was reduced in the cell somata from 2.5 ± 0.63 in control to 0.04 ± 0.03 in ryanodine (100 µM; P = 0.014, n = 5, FOV = 5). (I) In the cell processes, the number of Ca 2+ firing sites per FOV was reduced from 70 ± 13.12 in control to 2.8 ± 2.33 in ryanodine (100 µM; P = 0.011, n = 5, FOV = 5). (J) PTCL area/frame was reduced in the cell processes from 169.1 ± 59.46 µm 2 in control to 1.3 ± 1.2 µm 2 in ryanodine (100 µM; P = 0.046, n = 5, FOV = 5). (K) PTCL count/frame was reduced in the cell processes from 4.57 ± 1.59 in control to 0.045 ± 0.043 in ryanodine (100 µM; P = 0.014, n = 5, FOV = 5). *, P
    Figure Legend Snippet: The effect of ryanodine on Ca 2+ transients in ICC-MY. (A and B) Representative heat map showing the summated PTCLs for an entire 60× magnification recording of ICC-MY in control and ryanodine (100 µM) showing the inhibitory effects of ryanodine (A), as indicated in the occurrence map of individually color-coded Ca 2+ firing sites in the ICC-MY network (B). (C) Traces of PTCL activity (PTCL area [dark blue] and PTCL count [brown]) in control and ryanodine. (D and E) Histogram showing the probability (%) that an individual Ca 2+ firing site in the ICC-MY cell somata and cell processes in E will fire during a CTC cycle in control conditions (black bars) and ryanodine (100 µM; red bars; n = 5, FOV = 5). (F) The number of Ca 2+ firing sites per cell soma was reduced from 6.28 ± 1.47 in control to 0.94 ± 0.81 in the presence of ryanodine (100 µM; P = 0.01, n = 5, FOV = 5). (G) PTCL area/frame was reduced in the cell somata from 62.19 ± 6.34 µm 2 in control to 0.3 ± 0.2 µm 2 in ryanodine (100 µM; P = 0.002, n = 5, FOV = 5). (H) PTCL count/frame was reduced in the cell somata from 2.5 ± 0.63 in control to 0.04 ± 0.03 in ryanodine (100 µM; P = 0.014, n = 5, FOV = 5). (I) In the cell processes, the number of Ca 2+ firing sites per FOV was reduced from 70 ± 13.12 in control to 2.8 ± 2.33 in ryanodine (100 µM; P = 0.011, n = 5, FOV = 5). (J) PTCL area/frame was reduced in the cell processes from 169.1 ± 59.46 µm 2 in control to 1.3 ± 1.2 µm 2 in ryanodine (100 µM; P = 0.046, n = 5, FOV = 5). (K) PTCL count/frame was reduced in the cell processes from 4.57 ± 1.59 in control to 0.045 ± 0.043 in ryanodine (100 µM; P = 0.014, n = 5, FOV = 5). *, P

    Techniques Used: Immunocytochemistry, Activity Assay

    34) Product Images from "Enhancement of dendritic persistent Na+ currents by mGluR5 leads to an advancement of spike timing with an increase in temporal precision"

    Article Title: Enhancement of dendritic persistent Na+ currents by mGluR5 leads to an advancement of spike timing with an increase in temporal precision

    Journal: Molecular Brain

    doi: 10.1186/s13041-018-0410-7

    DHPG-induced increase of intrinsic excitability requires intracellular Ca 2+ . a Left, representative traces demonstrate the effects of DHPG on AP firing frequency and afterpotentials at 120 pA current injections when ryanodine (20 μM) was added to the pipette solutions. Right, FI curve summarizes the effects of DHPG on AP firing frequency in response to square pulse injections ( n = 4). b – d Summary plots of the FI curve under the conditions indicated ( b , n = 4, 5 mM nicotinamide; c , n = 5, 100 μM 8-NH 2 -cADPR; d , n = 5, 3 μM calmidazolium). e Bar graphs summarize the effects of DHPG on afterpotentials following 160 pA square current injections under different conditions indicated (n.s., P > 0.05, * P
    Figure Legend Snippet: DHPG-induced increase of intrinsic excitability requires intracellular Ca 2+ . a Left, representative traces demonstrate the effects of DHPG on AP firing frequency and afterpotentials at 120 pA current injections when ryanodine (20 μM) was added to the pipette solutions. Right, FI curve summarizes the effects of DHPG on AP firing frequency in response to square pulse injections ( n = 4). b – d Summary plots of the FI curve under the conditions indicated ( b , n = 4, 5 mM nicotinamide; c , n = 5, 100 μM 8-NH 2 -cADPR; d , n = 5, 3 μM calmidazolium). e Bar graphs summarize the effects of DHPG on afterpotentials following 160 pA square current injections under different conditions indicated (n.s., P > 0.05, * P

    Techniques Used: Transferring

    35) Product Images from "Calcium-permeable AMPA receptors mediate timing-dependent LTP elicited by 6 coincident action potentials at Schaffer collateral-CA1 synapses"

    Article Title: Calcium-permeable AMPA receptors mediate timing-dependent LTP elicited by 6 coincident action potentials at Schaffer collateral-CA1 synapses

    Journal: bioRxiv

    doi: 10.1101/719633

    Contribution of group I mGluRs, IP3 receptors and ryanodine receptor-dependent calcium release from internal stores to 6 x 1:4 t-LTP. A) Inclusion of 10 mM BAPTA in the pipette solution and equilibration with the cell interior for 30 min before t-LTP induction (open circles) prevented t-LTP induced by 6 x 1:4 stimulation compared to identically treated (i.e. t-LTP induction 30 min after breaking the patch) control cells (closed circles; Control: n=5 / N=5, BAPTA i : n=6 / N=4), indicating the necessity of postsynaptic calcium elevation to induce t-LTP. The inset depicts the loading of the cell with BAPTA. B) T-LTP induced with the 6 x 1:4 protocol was neither affected by bath application of the mGluR1 antagonist YM-298198 (1 µM; ACSF: n=7 / N=5, YM-298198: n=6 / N=3), nor by the mGluR5 antagonist MPEP (10 µM, n=6 / N=4). C) However, co-application of both antagonists (YM-298198 and MPEP; ACSF n=12 / N=8, YM-MPEP n=12 / N=5) significantly reduced synaptic potentiation. D) Inhibition of IP3 receptors by 100 µM 2-APB (in 0.05% DMSO) completely blocked 6 x 1:4 t-LTP (DMSO n=10/ N=5; 2-APB n=7/ N=3). E) Wash in of 100 µM ryanodine into the postsynaptic neuron via the patch pipette inhibited t-LTP induced by 6 x 1:4 stimulation (DMSO n= 9 / N= 4; Ryanodine n= 14 / N= 5). Average time course of potentiation and mean (± SEM) magnitude of t-LTP are shown for the respective experiments.
    Figure Legend Snippet: Contribution of group I mGluRs, IP3 receptors and ryanodine receptor-dependent calcium release from internal stores to 6 x 1:4 t-LTP. A) Inclusion of 10 mM BAPTA in the pipette solution and equilibration with the cell interior for 30 min before t-LTP induction (open circles) prevented t-LTP induced by 6 x 1:4 stimulation compared to identically treated (i.e. t-LTP induction 30 min after breaking the patch) control cells (closed circles; Control: n=5 / N=5, BAPTA i : n=6 / N=4), indicating the necessity of postsynaptic calcium elevation to induce t-LTP. The inset depicts the loading of the cell with BAPTA. B) T-LTP induced with the 6 x 1:4 protocol was neither affected by bath application of the mGluR1 antagonist YM-298198 (1 µM; ACSF: n=7 / N=5, YM-298198: n=6 / N=3), nor by the mGluR5 antagonist MPEP (10 µM, n=6 / N=4). C) However, co-application of both antagonists (YM-298198 and MPEP; ACSF n=12 / N=8, YM-MPEP n=12 / N=5) significantly reduced synaptic potentiation. D) Inhibition of IP3 receptors by 100 µM 2-APB (in 0.05% DMSO) completely blocked 6 x 1:4 t-LTP (DMSO n=10/ N=5; 2-APB n=7/ N=3). E) Wash in of 100 µM ryanodine into the postsynaptic neuron via the patch pipette inhibited t-LTP induced by 6 x 1:4 stimulation (DMSO n= 9 / N= 4; Ryanodine n= 14 / N= 5). Average time course of potentiation and mean (± SEM) magnitude of t-LTP are shown for the respective experiments.

    Techniques Used: Transferring, Inhibition

    Suggested cellular signaling mechanisms involved in low repeat t-LTP at Schaffer collateral-CA1 synapses. Summary of induction, signaling, and expression mechanisms involved in low repeat canonical (i.e. 6 x 1:1 t-LTP) and burst (i.e. 6 x 1:4) t-LTP protocols in CA1 pyramidal neurons. A) Synaptic mechanisms involved in the presynaptically expressed 6 x 1:1 t-LTP. T-LTP induction depends on postsynaptic NMDAR and L-type VGCC mediated Ca 2+ influx (1.). Insertion of cp-AMPARs into the postsynaptic membrane might be regulated by D1/D2 signaling (2.) and could account for the combined D1/D2 receptor dependence of 6 x 1:1 t-LTP. Ongoing low frequency test stimulation after induction of t-LTP leads to sustained Ca 2+ elevations through postsynaptic cp-AMPARs (3.). The resulting prolonged postsynaptic Ca 2+ elevation leads via a yet unidentified retrograde messenger to increased presynaptic efficacy (4.). An additional presynaptic contribution of D1/D2 signaling to enhanced presynaptic glutamatergic function is possible. B) The postsynaptically expressed 6 x 1:4 t-LTP does neither require postsynaptic NMDAR nor L-type VGCC activation for induction. It rather depends on calcium release from postsynaptic internal stores mediated by mGluR 1,5 -dependent activation of IP3 receptors in the ER (1.). This initial postsynaptic Ca 2+ rise is amplified by Ca 2+ dependent Ca 2+ release via Ryanodine receptors (RyRs; 2.). Moreover, the 6 x 1:4 t-LTP depends (like 6 x 1:1 t-LTP) on the activation of cp-AMPARs. Intact D2 receptor signaling is mandatory to observe 6 x 1:4 t-LTP and might be involved in recruiting cp-AMPARs to the postsynaptic membrane (3.) for sustained Ca 2+ influx during ongoing low frequency synaptic stimulation after t-LTP induction (4.). The resulting prolonged postsynaptic Ca 2+ elevation initiated by mGluRs, RyRs, and cp-AMPARs leads to postsynaptic expression of 6 x 1:4 t-LTP by insertion of new GluA1 and GluA2-containing AMPARs into the postsynaptic membrane (5.)
    Figure Legend Snippet: Suggested cellular signaling mechanisms involved in low repeat t-LTP at Schaffer collateral-CA1 synapses. Summary of induction, signaling, and expression mechanisms involved in low repeat canonical (i.e. 6 x 1:1 t-LTP) and burst (i.e. 6 x 1:4) t-LTP protocols in CA1 pyramidal neurons. A) Synaptic mechanisms involved in the presynaptically expressed 6 x 1:1 t-LTP. T-LTP induction depends on postsynaptic NMDAR and L-type VGCC mediated Ca 2+ influx (1.). Insertion of cp-AMPARs into the postsynaptic membrane might be regulated by D1/D2 signaling (2.) and could account for the combined D1/D2 receptor dependence of 6 x 1:1 t-LTP. Ongoing low frequency test stimulation after induction of t-LTP leads to sustained Ca 2+ elevations through postsynaptic cp-AMPARs (3.). The resulting prolonged postsynaptic Ca 2+ elevation leads via a yet unidentified retrograde messenger to increased presynaptic efficacy (4.). An additional presynaptic contribution of D1/D2 signaling to enhanced presynaptic glutamatergic function is possible. B) The postsynaptically expressed 6 x 1:4 t-LTP does neither require postsynaptic NMDAR nor L-type VGCC activation for induction. It rather depends on calcium release from postsynaptic internal stores mediated by mGluR 1,5 -dependent activation of IP3 receptors in the ER (1.). This initial postsynaptic Ca 2+ rise is amplified by Ca 2+ dependent Ca 2+ release via Ryanodine receptors (RyRs; 2.). Moreover, the 6 x 1:4 t-LTP depends (like 6 x 1:1 t-LTP) on the activation of cp-AMPARs. Intact D2 receptor signaling is mandatory to observe 6 x 1:4 t-LTP and might be involved in recruiting cp-AMPARs to the postsynaptic membrane (3.) for sustained Ca 2+ influx during ongoing low frequency synaptic stimulation after t-LTP induction (4.). The resulting prolonged postsynaptic Ca 2+ elevation initiated by mGluRs, RyRs, and cp-AMPARs leads to postsynaptic expression of 6 x 1:4 t-LTP by insertion of new GluA1 and GluA2-containing AMPARs into the postsynaptic membrane (5.)

    Techniques Used: Expressing, Activation Assay, Amplification

    36) Product Images from "Interactions of Dichlorodiphenyltrichloroethane (DDT) and Dichlorodiphenyldichloroethylene (DDE) With Skeletal Muscle Ryanodine Receptor Type 1"

    Article Title: Interactions of Dichlorodiphenyltrichloroethane (DDT) and Dichlorodiphenyldichloroethylene (DDE) With Skeletal Muscle Ryanodine Receptor Type 1

    Journal: Toxicological Sciences

    doi: 10.1093/toxsci/kfz120

    Both of the dichlorodiphenyltrichloroethane (DDT) and the dichlorodiphenyldichloroethylene (DDE) congeners potentiate Ca 2+ release from sarcoplasmic reticulum (SR) stores through direct interaction with ryanodine receptor type 1 (RyR1). (A), Representative traces detailing the macroscopic Ca 2+ efflux assay: rabbit junctional sarcoplasmic reticulum (JSR) vesicles were actively loaded with Ca 2+ to near maximal capacity and then exposed to either 0.1% DMSO (v/v) vehicle control, 10 µM o , p ′ - DDT, 10 µM p , p ′-DDT, 10 µM o , p ′-DDE, or 10 µM p , p ′-DDE. Following DMSO or DDx addition, 2 µM ruthenium red, a ryanodine receptor blocker, was added to determine if DDxmediated Ca 2+ release by direct interaction with RyR1. Lastly, 50 µM cyclopiazonic acid, a sarco/endoplasmic reticulum Ca 2+ -ATPase inhibitor, was added to prevent Ca 2+ reuptake to allow for calibration of Ca 2+ . (B) The initial 60 s of the Ca 2+ release trace for DMSO vehicle control (square trace), o , p ′ - DDT (triangle trace), p , p ′-DDT (inverted triangle trace), o , p ′-DDE (circle trace), or p , p ′-DDE (square trace) was assessed with a linear regression to determine Ca 2+ release rate (nmole Ca 2+ /s/mg JSR). (C), o , p ′-DDE (37-fold) and p , p ′-DDE (13-fold) significantly increased the rate of Ca 2+ efflux from Ca 2+ -loaded RyR1 SR vesicles compared with baseline leak rate, whereas both DDT congeners triggered a 6.5-fold difference. Three independent measurements replicated 3 times ( n = 3) from different rabbit JSR preparations under identical condition were summarized and plotted (*** p
    Figure Legend Snippet: Both of the dichlorodiphenyltrichloroethane (DDT) and the dichlorodiphenyldichloroethylene (DDE) congeners potentiate Ca 2+ release from sarcoplasmic reticulum (SR) stores through direct interaction with ryanodine receptor type 1 (RyR1). (A), Representative traces detailing the macroscopic Ca 2+ efflux assay: rabbit junctional sarcoplasmic reticulum (JSR) vesicles were actively loaded with Ca 2+ to near maximal capacity and then exposed to either 0.1% DMSO (v/v) vehicle control, 10 µM o , p ′ - DDT, 10 µM p , p ′-DDT, 10 µM o , p ′-DDE, or 10 µM p , p ′-DDE. Following DMSO or DDx addition, 2 µM ruthenium red, a ryanodine receptor blocker, was added to determine if DDxmediated Ca 2+ release by direct interaction with RyR1. Lastly, 50 µM cyclopiazonic acid, a sarco/endoplasmic reticulum Ca 2+ -ATPase inhibitor, was added to prevent Ca 2+ reuptake to allow for calibration of Ca 2+ . (B) The initial 60 s of the Ca 2+ release trace for DMSO vehicle control (square trace), o , p ′ - DDT (triangle trace), p , p ′-DDT (inverted triangle trace), o , p ′-DDE (circle trace), or p , p ′-DDE (square trace) was assessed with a linear regression to determine Ca 2+ release rate (nmole Ca 2+ /s/mg JSR). (C), o , p ′-DDE (37-fold) and p , p ′-DDE (13-fold) significantly increased the rate of Ca 2+ efflux from Ca 2+ -loaded RyR1 SR vesicles compared with baseline leak rate, whereas both DDT congeners triggered a 6.5-fold difference. Three independent measurements replicated 3 times ( n = 3) from different rabbit JSR preparations under identical condition were summarized and plotted (*** p

    Techniques Used:

    Pretreatment of HEK-RyR1 cells with either dichlorodiphenyltrichloroethane (DDT) or dichlorodiphenyldichloroethylene (DDE) congener for 12-h facilitated a biphasic effect on ryanodine receptor type 1 (RyR1)-sensitization to caffeine response. (A), Ca 2+ -transient response mediated by addition of 100 µM caffeine to HEK-RyR1 cells pretreated with 0.1–10 µM o , p ′-DDT (triangle trace), p , p ′-DDT (inverted triangle traces), o , p ′ - DDE (circle traces), or p , p ′-DDE (square traces) or 0.1% DMSO (v/v) vehicle control. The (B) amplitude and (C) under the curve (AUC) of the caffeine response were quantified and plotted for each concentration of DDT or DDE. Experiments were performed in triplicate and repeated 4 times ( n = 4).
    Figure Legend Snippet: Pretreatment of HEK-RyR1 cells with either dichlorodiphenyltrichloroethane (DDT) or dichlorodiphenyldichloroethylene (DDE) congener for 12-h facilitated a biphasic effect on ryanodine receptor type 1 (RyR1)-sensitization to caffeine response. (A), Ca 2+ -transient response mediated by addition of 100 µM caffeine to HEK-RyR1 cells pretreated with 0.1–10 µM o , p ′-DDT (triangle trace), p , p ′-DDT (inverted triangle traces), o , p ′ - DDE (circle traces), or p , p ′-DDE (square traces) or 0.1% DMSO (v/v) vehicle control. The (B) amplitude and (C) under the curve (AUC) of the caffeine response were quantified and plotted for each concentration of DDT or DDE. Experiments were performed in triplicate and repeated 4 times ( n = 4).

    Techniques Used: Concentration Assay

    Dichlorodiphenyltrichloroethane (DDT) and dichlorodiphenyldichloroethylene (DDE) require time to permeate HEK-RyR1 cells to sensitize ryanodine receptor type 1 (RyR1) receptors. (A and B), Fluo-4 fluorescence emission traces showing Ca 2+ -transient responses of HEK-RyR1 cells and HEK-Null cells. (A), Although addition of 100 µM caffeine simultaneously with 10 µM o , p ′-DDT, p , p ′-DDT, o , p ′-DDE, p , p ′-DDE, or 0.1% DMSO (v/v) vehicle control does not cause Ca 2+ release from HEK-Null cells, it stimulates Ca 2+ release from HEK-RyR1 cells. The degree of RyR1-stimulation by the congeners showed no significant differences compared with DMSO vehicle control as assessed by (C) amplitude and (E) area under the curve (AUC) post-activation. (B), However, addition of 10 µM of either DDT or DDE congener 100 s before addition of 100 µM caffeine sensitized RyR1to the activating effect of 100 µM caffeine, increasing the (D) amplitude and (F) AUC post-stimulation, significantly compared with DMSO vehicle control with caffeine. Experiments were performed in triplicate and repeated 4 times ( n = 4). Statistical comparison of the effect of DDT and DDE congeners to DMSO vehicle control was performed with a one-way ANOVA with Dunnett post hoc test (* p
    Figure Legend Snippet: Dichlorodiphenyltrichloroethane (DDT) and dichlorodiphenyldichloroethylene (DDE) require time to permeate HEK-RyR1 cells to sensitize ryanodine receptor type 1 (RyR1) receptors. (A and B), Fluo-4 fluorescence emission traces showing Ca 2+ -transient responses of HEK-RyR1 cells and HEK-Null cells. (A), Although addition of 100 µM caffeine simultaneously with 10 µM o , p ′-DDT, p , p ′-DDT, o , p ′-DDE, p , p ′-DDE, or 0.1% DMSO (v/v) vehicle control does not cause Ca 2+ release from HEK-Null cells, it stimulates Ca 2+ release from HEK-RyR1 cells. The degree of RyR1-stimulation by the congeners showed no significant differences compared with DMSO vehicle control as assessed by (C) amplitude and (E) area under the curve (AUC) post-activation. (B), However, addition of 10 µM of either DDT or DDE congener 100 s before addition of 100 µM caffeine sensitized RyR1to the activating effect of 100 µM caffeine, increasing the (D) amplitude and (F) AUC post-stimulation, significantly compared with DMSO vehicle control with caffeine. Experiments were performed in triplicate and repeated 4 times ( n = 4). Statistical comparison of the effect of DDT and DDE congeners to DMSO vehicle control was performed with a one-way ANOVA with Dunnett post hoc test (* p

    Techniques Used: Fluorescence, Activation Assay

    Acute treatment with either dichlorodiphenyltrichloroethane (DDT) or dichlorodiphenyldichloroethylene (DDE) congeners failed to mediate Ca 2+ release from ryanodine receptor type 1 (RyR1)-expressing HEK293T cells (HEK-RyR1). (A and B), Fluo-4 fluorescence emission traces showing Ca 2+ -transient responses of HEK-RyR1 cells and wild-type HEK293T cells (HEK-Null) to treatment with various compounds. (A), Immediate addition of 10 µM o , p ′-DDT, 10 µM p , p ′-DDT, 10 µM o , p ′-DDE, 10 µM p , p ′-DDE, or 0.1% DMSO (v/v) vehicle control did not facilitate Ca 2+ release from HEK-RyR1 cells or HEK-Null cells; whereas, addition of 100 µM caffeine, an RyR1 agonist, stimulated Ca 2+ release from stores in HEK-RyR1 cells but not HEK-Null cells. (B), Treatment of HEK-Null cells with 10 µM thapsigargin, a noncompetitive inhibitor of SERCA, mediated Ca 2+ release from stores. Experiments were performed in sextuplicate and repeated 4 times ( n = 4).
    Figure Legend Snippet: Acute treatment with either dichlorodiphenyltrichloroethane (DDT) or dichlorodiphenyldichloroethylene (DDE) congeners failed to mediate Ca 2+ release from ryanodine receptor type 1 (RyR1)-expressing HEK293T cells (HEK-RyR1). (A and B), Fluo-4 fluorescence emission traces showing Ca 2+ -transient responses of HEK-RyR1 cells and wild-type HEK293T cells (HEK-Null) to treatment with various compounds. (A), Immediate addition of 10 µM o , p ′-DDT, 10 µM p , p ′-DDT, 10 µM o , p ′-DDE, 10 µM p , p ′-DDE, or 0.1% DMSO (v/v) vehicle control did not facilitate Ca 2+ release from HEK-RyR1 cells or HEK-Null cells; whereas, addition of 100 µM caffeine, an RyR1 agonist, stimulated Ca 2+ release from stores in HEK-RyR1 cells but not HEK-Null cells. (B), Treatment of HEK-Null cells with 10 µM thapsigargin, a noncompetitive inhibitor of SERCA, mediated Ca 2+ release from stores. Experiments were performed in sextuplicate and repeated 4 times ( n = 4).

    Techniques Used: Expressing, Fluorescence

    Neither FK506-binding protein (FKBP12) nor ryanodine receptor type 1 (RyR1) protein expression levels are different between preparations from female mice and preparations from male mice. (A), RyR1 microsomal preparations from individual female mice ( n = 7) and individual male mice ( n = 7) were assessed. (B), Densitometry analysis and a Student’s t -test confirmed no significant differences in protein expression levels of FKBP12, RyR1, or the ratio of RyR1 to FKBP12 between the 2 sexes.
    Figure Legend Snippet: Neither FK506-binding protein (FKBP12) nor ryanodine receptor type 1 (RyR1) protein expression levels are different between preparations from female mice and preparations from male mice. (A), RyR1 microsomal preparations from individual female mice ( n = 7) and individual male mice ( n = 7) were assessed. (B), Densitometry analysis and a Student’s t -test confirmed no significant differences in protein expression levels of FKBP12, RyR1, or the ratio of RyR1 to FKBP12 between the 2 sexes.

    Techniques Used: Binding Assay, Expressing, Mouse Assay

    37) Product Images from "Mechanisms Involved in Nicotinic Acetylcholine Receptor-Induced Neurotransmitter Release from Sympathetic Nerve Terminals in the Mouse Vas Deferens"

    Article Title: Mechanisms Involved in Nicotinic Acetylcholine Receptor-Induced Neurotransmitter Release from Sympathetic Nerve Terminals in the Mouse Vas Deferens

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0029209

    The effect of ryanodine on epibatidine-induced, electrically-evoked, and spontaneous neurotransmitter release. (A) A typical membrane potential trace showing the effects of epibatidine application following exposure to 10 µM ryanodine for 60 minutes. (B) A cumulative frequency plot of EID amplitude in the presence of ryanodine. There is a significant shift to smaller EID amplitudes in the presence of ryanodine (P
    Figure Legend Snippet: The effect of ryanodine on epibatidine-induced, electrically-evoked, and spontaneous neurotransmitter release. (A) A typical membrane potential trace showing the effects of epibatidine application following exposure to 10 µM ryanodine for 60 minutes. (B) A cumulative frequency plot of EID amplitude in the presence of ryanodine. There is a significant shift to smaller EID amplitudes in the presence of ryanodine (P

    Techniques Used:

    38) Product Images from "Oxidative Stress Induces Disruption of the Axon Initial Segment"

    Article Title: Oxidative Stress Induces Disruption of the Axon Initial Segment

    Journal: ASN NEURO

    doi: 10.1177/1759091417745426

    IP 3 -gated Ca 2+ stores are required for ROS/RNS-induced AIS disruption. AIS labeling (AnkG, red) was lost (white arrows) following exposure of cortical neurons (NeuN+, green) to 25 μM SIN-1 (b). This disruption was prevented by pretreatment with an IP 3 -receptor inhibitor ((d) and (f)) but not an inhibitor to ryanodine receptors ((c) and (e)). Asterisks without an associated bracket represent a significant difference from the SIN-1 untreated group (a, * p
    Figure Legend Snippet: IP 3 -gated Ca 2+ stores are required for ROS/RNS-induced AIS disruption. AIS labeling (AnkG, red) was lost (white arrows) following exposure of cortical neurons (NeuN+, green) to 25 μM SIN-1 (b). This disruption was prevented by pretreatment with an IP 3 -receptor inhibitor ((d) and (f)) but not an inhibitor to ryanodine receptors ((c) and (e)). Asterisks without an associated bracket represent a significant difference from the SIN-1 untreated group (a, * p

    Techniques Used: Labeling

    39) Product Images from "Anandamide potentiation of miniature spontaneous excitatory synaptic transmission is mediated via an IP3 pathway"

    Article Title: Anandamide potentiation of miniature spontaneous excitatory synaptic transmission is mediated via an IP3 pathway

    Journal: Neurochemistry international

    doi: 10.1016/j.neuint.2010.01.001

    Ryanodine receptor does not mediate the AEA-induced effect on mEPSCs. (A). Representative sweeps of mEPSCs in neurons treated with ryanodine (ryanodine receptor antagonist, 20 μM) in the absence or presence of AEA (5 μM) and washout. (B).
    Figure Legend Snippet: Ryanodine receptor does not mediate the AEA-induced effect on mEPSCs. (A). Representative sweeps of mEPSCs in neurons treated with ryanodine (ryanodine receptor antagonist, 20 μM) in the absence or presence of AEA (5 μM) and washout. (B).

    Techniques Used:

    40) Product Images from "Group I mGluRs Evoke K-ATP Current by Intracellular Ca2+ Mobilization in Rat Subthalamus Neurons"

    Article Title: Group I mGluRs Evoke K-ATP Current by Intracellular Ca2+ Mobilization in Rat Subthalamus Neurons

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    doi: 10.1124/jpet.112.201566

    Ryanodine- and InsP 3 -gated Ca 2+ release are required for DHPG-induced conductance increase. (A) I–V plots showing that bath application of ryanodine (10 µ M) significantly altered the DHPG I-V plot ( n = 5; P
    Figure Legend Snippet: Ryanodine- and InsP 3 -gated Ca 2+ release are required for DHPG-induced conductance increase. (A) I–V plots showing that bath application of ryanodine (10 µ M) significantly altered the DHPG I-V plot ( n = 5; P

    Techniques Used:

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    Article Title: Cerebellar Kainate Receptor-Mediated Facilitation of Glutamate Release Requires Ca2+-Calmodulin and PKA
    Article Snippet: Salts and general reagents were purchased from Sigma (St. Louis, MO, USA); GYKI 53655, D-AP5, NBQX, bicuculline, Rp-Br-cAMP, H-89, forskolin, philanthotoxin, ryanodine, thapsigargin, kainate, Pertussis toxin CMZ and W-7 were obtained from Tocris (Bristol, UK).

    Article Title: Ryanodine Receptor-Mediated Calcium Release Has a Key Role in Hippocampal LTD Induction
    Article Snippet: Stock solutions of ryanodine (Tocris, Bristol, UK) and caffeine (Sigma, St. Louis, MO, USA) were dissolved in water and stored in aliquots at −20°C before thawing and dilution to their final concentrations in artificial cerebrospinal fluid (ACSF) solution (in mM: 124 NaCl, 5 KCl, 1 MgCl2 , 2 CaCl2 , 1.25 NaH2 PO4 , 26 NaHCO3 , pH 7.4, 10 glucose).

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    Article Snippet: Ryanodine and GSK-429286 were from Tocris Bioscience.

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    Tocris drugs ryanodine
    Caffeine induces an NMDAR‐independent LTP that does not occlude the classical LTP evoked by high frequency stimulation. (A) Summary of time‐course of mean fEPSPs slope in basal conditions and following bath application of 100‐μM caffeine (5 min, bar) in control slices (filled circles, n = 12; N = 7) and in the presence of 50‐μM AP5 (empty circles, n = 7; N = 3). Traces inset in the plots represent fEPSPs averages recorded during periods indicated by corresponding numbers in the graph. (B) Summary data showing mean fEPSP slopes in hippocampal slices before (Basal) and after (5 and 60 min) application of caffeine in control condition ( n = 12; N = 7; same as (A)) and in the presence of 50‐μM AP5 ( n = 7; N = 3), 50‐μM picrotoxin (PTX, n = 7; N = 3) and 20‐μM <t>ryanodine</t> (Ryan, n = 7; N = 3). (C) Mean average of paired‐pulse facilitation (PPF) at interstimulus interval of 50 ms in basal condition and after perfusion with caffeine at different concentrations (100 μM: n = 12; N = 6; 500 μM: n = 14; N = 6; 5 mM: n = 7; N = 3). Insets of tracings in the plots represent fEPSPs averages recorded in basal conditions and after 5 and 60 min of 100‐μM caffeine perfusion. (D) Summary data showing LTP induced by 100‐μM caffeine (5 min, bar) following by high frequency stimulation of SC (HFS ↑) after 30 min (filled circles, n = 11; N = 4). Significant differences with respect to basal state were established with Student's t ‐test at * P
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    Caffeine induces an NMDAR‐independent LTP that does not occlude the classical LTP evoked by high frequency stimulation. (A) Summary of time‐course of mean fEPSPs slope in basal conditions and following bath application of 100‐μM caffeine (5 min, bar) in control slices (filled circles, n = 12; N = 7) and in the presence of 50‐μM AP5 (empty circles, n = 7; N = 3). Traces inset in the plots represent fEPSPs averages recorded during periods indicated by corresponding numbers in the graph. (B) Summary data showing mean fEPSP slopes in hippocampal slices before (Basal) and after (5 and 60 min) application of caffeine in control condition ( n = 12; N = 7; same as (A)) and in the presence of 50‐μM AP5 ( n = 7; N = 3), 50‐μM picrotoxin (PTX, n = 7; N = 3) and 20‐μM ryanodine (Ryan, n = 7; N = 3). (C) Mean average of paired‐pulse facilitation (PPF) at interstimulus interval of 50 ms in basal condition and after perfusion with caffeine at different concentrations (100 μM: n = 12; N = 6; 500 μM: n = 14; N = 6; 5 mM: n = 7; N = 3). Insets of tracings in the plots represent fEPSPs averages recorded in basal conditions and after 5 and 60 min of 100‐μM caffeine perfusion. (D) Summary data showing LTP induced by 100‐μM caffeine (5 min, bar) following by high frequency stimulation of SC (HFS ↑) after 30 min (filled circles, n = 11; N = 4). Significant differences with respect to basal state were established with Student's t ‐test at * P

    Journal: Addiction Biology

    Article Title: Caffeine‐mediated BDNF release regulates long‐term synaptic plasticity through activation of IRS2 signaling) Caffeine‐mediated BDNF release regulates long‐term synaptic plasticity through activation of IRS2 signaling

    doi: 10.1111/adb.12433

    Figure Lengend Snippet: Caffeine induces an NMDAR‐independent LTP that does not occlude the classical LTP evoked by high frequency stimulation. (A) Summary of time‐course of mean fEPSPs slope in basal conditions and following bath application of 100‐μM caffeine (5 min, bar) in control slices (filled circles, n = 12; N = 7) and in the presence of 50‐μM AP5 (empty circles, n = 7; N = 3). Traces inset in the plots represent fEPSPs averages recorded during periods indicated by corresponding numbers in the graph. (B) Summary data showing mean fEPSP slopes in hippocampal slices before (Basal) and after (5 and 60 min) application of caffeine in control condition ( n = 12; N = 7; same as (A)) and in the presence of 50‐μM AP5 ( n = 7; N = 3), 50‐μM picrotoxin (PTX, n = 7; N = 3) and 20‐μM ryanodine (Ryan, n = 7; N = 3). (C) Mean average of paired‐pulse facilitation (PPF) at interstimulus interval of 50 ms in basal condition and after perfusion with caffeine at different concentrations (100 μM: n = 12; N = 6; 500 μM: n = 14; N = 6; 5 mM: n = 7; N = 3). Insets of tracings in the plots represent fEPSPs averages recorded in basal conditions and after 5 and 60 min of 100‐μM caffeine perfusion. (D) Summary data showing LTP induced by 100‐μM caffeine (5 min, bar) following by high frequency stimulation of SC (HFS ↑) after 30 min (filled circles, n = 11; N = 4). Significant differences with respect to basal state were established with Student's t ‐test at * P

    Article Snippet: Drugs Ryanodine, 8‐Cyclopentyl‐1,3‐dipropylxanthine (DPCPX), thapsigargin, suramin, and D (−)‐2‐amino‐5‐phosphonopentanoic acid (AP5) were purchased from Tocris Cookson (Bristol, UK).

    Techniques: Mass Spectrometry

    Caffeine‐induced BDNF release and TrkB activation contribute to the maintenance of CAF LTP. (A) Effect of caffeine on BDNF release (percent respect to basal) following 100‐μM caffeine application (5 min) in control conditions ( n = 27; N = 4) and after treatment with tetrodotoxin (TTX, n = 12; N = 4) or ryanodine (Ryan, n = 15; N = 4). Significant differences were established at *** P

    Journal: Addiction Biology

    Article Title: Caffeine‐mediated BDNF release regulates long‐term synaptic plasticity through activation of IRS2 signaling) Caffeine‐mediated BDNF release regulates long‐term synaptic plasticity through activation of IRS2 signaling

    doi: 10.1111/adb.12433

    Figure Lengend Snippet: Caffeine‐induced BDNF release and TrkB activation contribute to the maintenance of CAF LTP. (A) Effect of caffeine on BDNF release (percent respect to basal) following 100‐μM caffeine application (5 min) in control conditions ( n = 27; N = 4) and after treatment with tetrodotoxin (TTX, n = 12; N = 4) or ryanodine (Ryan, n = 15; N = 4). Significant differences were established at *** P

    Article Snippet: Drugs Ryanodine, 8‐Cyclopentyl‐1,3‐dipropylxanthine (DPCPX), thapsigargin, suramin, and D (−)‐2‐amino‐5‐phosphonopentanoic acid (AP5) were purchased from Tocris Cookson (Bristol, UK).

    Techniques: Activation Assay

    LTD-induced phosphorylation of CaMKII and Synapsin I. Induction of LTD for 60 min in control (C) slices caused a significant increase in the phosphorylation levels of Synapsin I (left panels), CaMKII-α (center panels) and CaMKII-β (right panels) relative to the levels displayed by unstimulated slices (naïve). Slices pre-incubated for 1 h with 20 μM ryanodine (Rya) before applying the LTD induction protocol displayed significantly lower increments in the phosphorylation levels of Synapsin I and CaMKII-α, whereas the phosphorylation levels of CaMKII-β were not significantly different from the levels displayed by unstimulated slices. Values represent Mean ± SE ( n = 3). Statistical analysis was performed with one-way ANOVA, followed by Tukey’s post hoc test. * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Ryanodine Receptor-Mediated Calcium Release Has a Key Role in Hippocampal LTD Induction

    doi: 10.3389/fncel.2018.00403

    Figure Lengend Snippet: LTD-induced phosphorylation of CaMKII and Synapsin I. Induction of LTD for 60 min in control (C) slices caused a significant increase in the phosphorylation levels of Synapsin I (left panels), CaMKII-α (center panels) and CaMKII-β (right panels) relative to the levels displayed by unstimulated slices (naïve). Slices pre-incubated for 1 h with 20 μM ryanodine (Rya) before applying the LTD induction protocol displayed significantly lower increments in the phosphorylation levels of Synapsin I and CaMKII-α, whereas the phosphorylation levels of CaMKII-β were not significantly different from the levels displayed by unstimulated slices. Values represent Mean ± SE ( n = 3). Statistical analysis was performed with one-way ANOVA, followed by Tukey’s post hoc test. * p

    Article Snippet: Stock solutions of ryanodine (Tocris, Bristol, UK) and caffeine (Sigma, St. Louis, MO, USA) were dissolved in water and stored in aliquots at −20°C before thawing and dilution to their final concentrations in artificial cerebrospinal fluid (ACSF) solution (in mM: 124 NaCl, 5 KCl, 1 MgCl2 , 2 CaCl2 , 1.25 NaH2 PO4 , 26 NaHCO3 , pH 7.4, 10 glucose).

    Techniques: Incubation

    Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on fEPSP rise times, half-widths and decay constants tau measured in CA3–CA1 hippocampal synapses. Left panels: slices were treated with 1 μM ryanodine. (A) fEPSP rise times vs. stimulus intensity. (B) fEPSP half-widths vs. stimulus intensity; (C) fEPSP decay constants (tau) vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine for 60 min. (D) fEPSP rise times vs. stimulus intensity; (E) fEPSP half-widths vs. stimulus intensity; (F) fEPSP decay constants (tau) vs. stimulus intensity. Right panels: slices were treated for 15 min with 1 mM caffeine. (G) fEPSP rise times vs. stimulus intensity; (H) fEPSP half-widths vs. stimulus intensity; (I) fEPSP decay constants (tau) vs. stimulus intensity. Values represent Mean ± SE; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed Mann-Whitney test (* p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Ryanodine Receptor-Mediated Calcium Release Has a Key Role in Hippocampal LTD Induction

    doi: 10.3389/fncel.2018.00403

    Figure Lengend Snippet: Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on fEPSP rise times, half-widths and decay constants tau measured in CA3–CA1 hippocampal synapses. Left panels: slices were treated with 1 μM ryanodine. (A) fEPSP rise times vs. stimulus intensity. (B) fEPSP half-widths vs. stimulus intensity; (C) fEPSP decay constants (tau) vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine for 60 min. (D) fEPSP rise times vs. stimulus intensity; (E) fEPSP half-widths vs. stimulus intensity; (F) fEPSP decay constants (tau) vs. stimulus intensity. Right panels: slices were treated for 15 min with 1 mM caffeine. (G) fEPSP rise times vs. stimulus intensity; (H) fEPSP half-widths vs. stimulus intensity; (I) fEPSP decay constants (tau) vs. stimulus intensity. Values represent Mean ± SE; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed Mann-Whitney test (* p

    Article Snippet: Stock solutions of ryanodine (Tocris, Bristol, UK) and caffeine (Sigma, St. Louis, MO, USA) were dissolved in water and stored in aliquots at −20°C before thawing and dilution to their final concentrations in artificial cerebrospinal fluid (ACSF) solution (in mM: 124 NaCl, 5 KCl, 1 MgCl2 , 2 CaCl2 , 1.25 NaH2 PO4 , 26 NaHCO3 , pH 7.4, 10 glucose).

    Techniques: MANN-WHITNEY

    Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on paired-pulse (PP) responses measured in CA3–CA1 hippocampal synapses. (A) Representative fEPSP traces showing PP responses before and after addition of 1 μM ryanodine (left panels), 20 μM ryanodine (center panels) and 1 mM caffeine (right panels). (B) Effects of 1 μM ryanodine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (D) Effects of 1 mM caffeine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (C) Effects of 20 μM ryanodine applied for 60 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. Values represent Mean ± SE; (13, 4) for ryanodine-treated slices; (12, 3) for caffeine-treated slices and (16, 4) for the control. The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Ryanodine Receptor-Mediated Calcium Release Has a Key Role in Hippocampal LTD Induction

    doi: 10.3389/fncel.2018.00403

    Figure Lengend Snippet: Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on paired-pulse (PP) responses measured in CA3–CA1 hippocampal synapses. (A) Representative fEPSP traces showing PP responses before and after addition of 1 μM ryanodine (left panels), 20 μM ryanodine (center panels) and 1 mM caffeine (right panels). (B) Effects of 1 μM ryanodine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (D) Effects of 1 mM caffeine applied for 15 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. (C) Effects of 20 μM ryanodine applied for 60 min on PP facilitation. The graph illustrates the facilitation ratio vs. inter-stimulus intervals. Values represent Mean ± SE; (13, 4) for ryanodine-treated slices; (12, 3) for caffeine-treated slices and (16, 4) for the control. The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used.

    Article Snippet: Stock solutions of ryanodine (Tocris, Bristol, UK) and caffeine (Sigma, St. Louis, MO, USA) were dissolved in water and stored in aliquots at −20°C before thawing and dilution to their final concentrations in artificial cerebrospinal fluid (ACSF) solution (in mM: 124 NaCl, 5 KCl, 1 MgCl2 , 2 CaCl2 , 1.25 NaH2 PO4 , 26 NaHCO3 , pH 7.4, 10 glucose).

    Techniques:

    Stimulatory and inhibitory ryanodine concentrations and caffeine modify the long-term depression (LTD) response. (A) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the low frequency stimulation (LFS) protocol to control hippocampal slices (14, 4) or to slices treated with 1 μM ryanodine (14, 3) or 20 μM ryanodine (13, 4). Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min after applying the LFS protocol (trace 2) to control slices, or recorded in slices treated with 1 μM or 20 μM ryanodine are shown on top of the graph. Open symbols: control slices; gray symbols: slices treated with 1 μM ryanodine; black symbols: slices treated with 20 μM ryanodine. (B) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation. (C) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the LFS protocol to control hippocampal slices (15, 3) or to slices treated with 1 mM caffeine (12, 4). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min (trace 2) after applying the LFS protocol to control slices and to slices treated with 1 mM caffeine are shown on top of the graph. Open symbols: control slices; black symbols: slices treated with 1 mM caffeine. (D) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation of control or caffeine-treated slices. Values represent Mean ± SE. Statistical significance of values was assessed by Mann-Whitney test (* p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Ryanodine Receptor-Mediated Calcium Release Has a Key Role in Hippocampal LTD Induction

    doi: 10.3389/fncel.2018.00403

    Figure Lengend Snippet: Stimulatory and inhibitory ryanodine concentrations and caffeine modify the long-term depression (LTD) response. (A) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the low frequency stimulation (LFS) protocol to control hippocampal slices (14, 4) or to slices treated with 1 μM ryanodine (14, 3) or 20 μM ryanodine (13, 4). Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min after applying the LFS protocol (trace 2) to control slices, or recorded in slices treated with 1 μM or 20 μM ryanodine are shown on top of the graph. Open symbols: control slices; gray symbols: slices treated with 1 μM ryanodine; black symbols: slices treated with 20 μM ryanodine. (B) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation. (C) Time course of fEPSP slopes recorded (CA3–CA1) before and after application of the LFS protocol to control hippocampal slices (15, 3) or to slices treated with 1 mM caffeine (12, 4). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Representative fEPSP traces recorded 1–5 min before (trace 1) and 60 min (trace 2) after applying the LFS protocol to control slices and to slices treated with 1 mM caffeine are shown on top of the graph. Open symbols: control slices; black symbols: slices treated with 1 mM caffeine. (D) Average magnitudes of fEPSP slopes recorded during the last 10 min after stimulation of control or caffeine-treated slices. Values represent Mean ± SE. Statistical significance of values was assessed by Mann-Whitney test (* p

    Article Snippet: Stock solutions of ryanodine (Tocris, Bristol, UK) and caffeine (Sigma, St. Louis, MO, USA) were dissolved in water and stored in aliquots at −20°C before thawing and dilution to their final concentrations in artificial cerebrospinal fluid (ACSF) solution (in mM: 124 NaCl, 5 KCl, 1 MgCl2 , 2 CaCl2 , 1.25 NaH2 PO4 , 26 NaHCO3 , pH 7.4, 10 glucose).

    Techniques: MANN-WHITNEY

    Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on basal synaptic transmission in the CA3–CA1 hippocampal synapse. Left panels: slices were treated with 1 μM ryanodine; representative field excitatory postsynaptic potential (fEPSP) traces registered before and 15 min after ryanodine addition are illustrated on top of the panels. (A) Fiber volley (FV) amplitude vs. stimulus intensity; (B) amplitude vs. stimulus intensity; (C) fEPSP slopes vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine; representative fEPSP traces registered before and 60 min after ryanodine addition are illustrated on top. (D) FV amplitude vs. stimulus intensity; (E) fEPSP amplitude vs. stimulus intensity; (F) fEPSP slopes vs. stimulus intensity. Right Panels: slices were treated with 1 mM caffeine; representative fEPSP traces registered before and 15 min after caffeine addition are illustrated on top. (G) FV amplitude vs. stimulus intensity; (H) fEPSP amplitude vs. stimulus intensity; (I) fEPSP slopes vs. stimulus intensity. Values represent Mean ± SEM; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed by Mann-Whitney test ( p > 0.05 in all cases).

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Ryanodine Receptor-Mediated Calcium Release Has a Key Role in Hippocampal LTD Induction

    doi: 10.3389/fncel.2018.00403

    Figure Lengend Snippet: Effects of stimulatory and inhibitory ryanodine concentrations and of caffeine on basal synaptic transmission in the CA3–CA1 hippocampal synapse. Left panels: slices were treated with 1 μM ryanodine; representative field excitatory postsynaptic potential (fEPSP) traces registered before and 15 min after ryanodine addition are illustrated on top of the panels. (A) Fiber volley (FV) amplitude vs. stimulus intensity; (B) amplitude vs. stimulus intensity; (C) fEPSP slopes vs. stimulus intensity. Central panels: slices were treated with 20 μM ryanodine; representative fEPSP traces registered before and 60 min after ryanodine addition are illustrated on top. (D) FV amplitude vs. stimulus intensity; (E) fEPSP amplitude vs. stimulus intensity; (F) fEPSP slopes vs. stimulus intensity. Right Panels: slices were treated with 1 mM caffeine; representative fEPSP traces registered before and 15 min after caffeine addition are illustrated on top. (G) FV amplitude vs. stimulus intensity; (H) fEPSP amplitude vs. stimulus intensity; (I) fEPSP slopes vs. stimulus intensity. Values represent Mean ± SEM; (12, 3). The first number in parentheses indicates the number of hippocampal slices and the second the number of animals used. Statistical significance of values was assessed by Mann-Whitney test ( p > 0.05 in all cases).

    Article Snippet: Stock solutions of ryanodine (Tocris, Bristol, UK) and caffeine (Sigma, St. Louis, MO, USA) were dissolved in water and stored in aliquots at −20°C before thawing and dilution to their final concentrations in artificial cerebrospinal fluid (ACSF) solution (in mM: 124 NaCl, 5 KCl, 1 MgCl2 , 2 CaCl2 , 1.25 NaH2 PO4 , 26 NaHCO3 , pH 7.4, 10 glucose).

    Techniques: Transmission Assay, MANN-WHITNEY

    Effect of 30 mM K + on whole-cell spatial and temporal Ca 2+ signaling characteristics recorded in situ. A : baseline-subtracted fractional fluorescence (F/F o ) traces for Fluo-4 fluorescence are shown for individual pulmonary arterial myocytes from a normoxic adult sheep. Recordings were made in the absence (control) or presence of 30 mM K + (30K) with or without 10 μM ryanodine (30K RY). Bars indicate means ± SE for normoxic (open) and long-term hypoxic (solid) conditions. B, D, and F : number of myocytes with Ca 2+ responses each minute in a 1,000 μm 2 area. C, E, and G : frequency of Ca 2+ events *,§,† P

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Maternal high-altitude hypoxia and suppression of ryanodine receptor-mediated Ca2+ sparks in fetal sheep pulmonary arterial myocytes

    doi: 10.1152/ajplung.00009.2012

    Figure Lengend Snippet: Effect of 30 mM K + on whole-cell spatial and temporal Ca 2+ signaling characteristics recorded in situ. A : baseline-subtracted fractional fluorescence (F/F o ) traces for Fluo-4 fluorescence are shown for individual pulmonary arterial myocytes from a normoxic adult sheep. Recordings were made in the absence (control) or presence of 30 mM K + (30K) with or without 10 μM ryanodine (30K RY). Bars indicate means ± SE for normoxic (open) and long-term hypoxic (solid) conditions. B, D, and F : number of myocytes with Ca 2+ responses each minute in a 1,000 μm 2 area. C, E, and G : frequency of Ca 2+ events *,§,† P

    Article Snippet: Ryanodine and dantrolene were from Tocris (Ellisville, MO).

    Techniques: In Situ, Fluorescence

    Effect of emodepside (1 µM) on membrane spiking. (A) Representative current-clamp trace showing the spiking induced by ryanodine (1 µM) and the inhibitory effect of emodepside on the spikes. (B) Bar chart (mean ± SEM) of the effects

    Journal: British Journal of Pharmacology

    Article Title: On the mode of action of emodepside: slow effects on membrane potential and voltage-activated currents in Ascaris suum

    doi: 10.1111/j.1476-5381.2011.01428.x

    Figure Lengend Snippet: Effect of emodepside (1 µM) on membrane spiking. (A) Representative current-clamp trace showing the spiking induced by ryanodine (1 µM) and the inhibitory effect of emodepside on the spikes. (B) Bar chart (mean ± SEM) of the effects

    Article Snippet: Ryanodine, staurosporine, iberiotoxin and PMA were obtained from Tocris Biosciences (Ellisville, MO, USA); 2 mM stock ryanodine was made every week.

    Techniques: