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    Structured Review

    Alomone Labs ttx
    Effects of veratridine on human sperm motility in the presence of tetrodotoxin, <t>A-803467</t> or ab-66743. The effects of veratridine (10 μM) after different incubation times were analyzed in the presence of (A) the VGSC inhibitor tetrodotoxin <t>(TTX)</t> (10 nM) (B) TTX (10 μM), (C) the selective Na v 1.8 antagonist A-803467 (10 μM), (D) the Na v 1.8 antibody ab-66743 (dilution 1:50) or the corresponding solvent. Bars are means with SEM of 6-8 different experiments and represent percentage changes in progressive motility (grade A+B sperm) relative to the value observed at the same time in the respective solvent-treated paired controls. * P
    Ttx, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ttx/product/Alomone Labs
    Average 96 stars, based on 39 article reviews
    Price from $9.99 to $1999.99
    ttx - by Bioz Stars, 2022-08
    96/100 stars

    Images

    1) Product Images from "The Voltage-Gated Sodium Channel Nav1.8 Is Expressed in Human Sperm"

    Article Title: The Voltage-Gated Sodium Channel Nav1.8 Is Expressed in Human Sperm

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0076084

    Effects of veratridine on human sperm motility in the presence of tetrodotoxin, A-803467 or ab-66743. The effects of veratridine (10 μM) after different incubation times were analyzed in the presence of (A) the VGSC inhibitor tetrodotoxin (TTX) (10 nM) (B) TTX (10 μM), (C) the selective Na v 1.8 antagonist A-803467 (10 μM), (D) the Na v 1.8 antibody ab-66743 (dilution 1:50) or the corresponding solvent. Bars are means with SEM of 6-8 different experiments and represent percentage changes in progressive motility (grade A+B sperm) relative to the value observed at the same time in the respective solvent-treated paired controls. * P
    Figure Legend Snippet: Effects of veratridine on human sperm motility in the presence of tetrodotoxin, A-803467 or ab-66743. The effects of veratridine (10 μM) after different incubation times were analyzed in the presence of (A) the VGSC inhibitor tetrodotoxin (TTX) (10 nM) (B) TTX (10 μM), (C) the selective Na v 1.8 antagonist A-803467 (10 μM), (D) the Na v 1.8 antibody ab-66743 (dilution 1:50) or the corresponding solvent. Bars are means with SEM of 6-8 different experiments and represent percentage changes in progressive motility (grade A+B sperm) relative to the value observed at the same time in the respective solvent-treated paired controls. * P

    Techniques Used: Incubation

    2) Product Images from "Neuroepithelial progenitors generate and propagate non-neuronal action potentials across the spinal cord"

    Article Title: Neuroepithelial progenitors generate and propagate non-neuronal action potentials across the spinal cord

    Journal: bioRxiv

    doi: 10.1101/2020.05.23.111955

    Floor-plate biphasic action potentials are triggered by the activation of nicotinic acetylcholine receptors in response to acetylcholine released by motoneurons. a, Example of recurrent spontaneous floor-plate action potentials blocked after addition of the nicotinic acetylcholine receptor (nAChR) antagonists: Mecamylamine (50 μM) and d-Tubocurarine (5 μM). b, Example of current-clamp recording showing floor-plate action potential evoked by electrical stimulation in control condition (black trace) and after addition of the nAChR antagonists (red trace). c, Example of current-clamp recording showing floor-plate action potential evoked in control condition (black trace) and after addition of antagonists against ionotropic receptor for GABA (Gabazine 3 μM) and glutamate (DL-APV 200 μM and CNQX 20 μM). d, Example of current-clamp recording showing that floor-plate action potential can be evoked by local application of 30 μM acetylcholine (left trace), even in the presence of TTX (1 μM) and antagonists to AMPA/Kainate glutamate receptor (CNQX 10 μM), NMDA glutamate receptor (DL-APV 200 μM), GABA A receptor (Gabazine 3 μM) and glycine receptor (strychnine 1 μM) (center trace). Floor-plate action potential evoked by acetylcholine were blocked by the addition of nAChR antagonists (right trace). Note that the addition of TTX inhibited the fast component of the biphasic action potential (see Supplementary Figure 2 ). e, Confocal image of a coronal section from a ChAT:ChR2-YFP mouse embryo at E12.5 showing the expression of Channelrhodopsin2-YFP fusion protein (in green) in cholinergic motoneurons located in the ventro-dorsal horns and labelled with the vesicular acetylcholine transporter vAChT (in red). All cell nuclei were labelled using DAPI (in blue). f, Example of current-clamp recording from a ChAT:ChR2-YFP + motoneuron showing an action potential triggered by the opening of Channelrhodopsin 2 in response to blue light stimulation (470 nm). g, Example of current-clamp recording from a floor-plate cell recorded in a ChAT:ChR2-YFP + fetal spinal cord showing how blue light stimulation could evoke a slow cholinergic depolarization and trigger a biphasic action potential that were blocked by the addition of nAChR antagonists.
    Figure Legend Snippet: Floor-plate biphasic action potentials are triggered by the activation of nicotinic acetylcholine receptors in response to acetylcholine released by motoneurons. a, Example of recurrent spontaneous floor-plate action potentials blocked after addition of the nicotinic acetylcholine receptor (nAChR) antagonists: Mecamylamine (50 μM) and d-Tubocurarine (5 μM). b, Example of current-clamp recording showing floor-plate action potential evoked by electrical stimulation in control condition (black trace) and after addition of the nAChR antagonists (red trace). c, Example of current-clamp recording showing floor-plate action potential evoked in control condition (black trace) and after addition of antagonists against ionotropic receptor for GABA (Gabazine 3 μM) and glutamate (DL-APV 200 μM and CNQX 20 μM). d, Example of current-clamp recording showing that floor-plate action potential can be evoked by local application of 30 μM acetylcholine (left trace), even in the presence of TTX (1 μM) and antagonists to AMPA/Kainate glutamate receptor (CNQX 10 μM), NMDA glutamate receptor (DL-APV 200 μM), GABA A receptor (Gabazine 3 μM) and glycine receptor (strychnine 1 μM) (center trace). Floor-plate action potential evoked by acetylcholine were blocked by the addition of nAChR antagonists (right trace). Note that the addition of TTX inhibited the fast component of the biphasic action potential (see Supplementary Figure 2 ). e, Confocal image of a coronal section from a ChAT:ChR2-YFP mouse embryo at E12.5 showing the expression of Channelrhodopsin2-YFP fusion protein (in green) in cholinergic motoneurons located in the ventro-dorsal horns and labelled with the vesicular acetylcholine transporter vAChT (in red). All cell nuclei were labelled using DAPI (in blue). f, Example of current-clamp recording from a ChAT:ChR2-YFP + motoneuron showing an action potential triggered by the opening of Channelrhodopsin 2 in response to blue light stimulation (470 nm). g, Example of current-clamp recording from a floor-plate cell recorded in a ChAT:ChR2-YFP + fetal spinal cord showing how blue light stimulation could evoke a slow cholinergic depolarization and trigger a biphasic action potential that were blocked by the addition of nAChR antagonists.

    Techniques Used: Activation Assay, Expressing

    3) Product Images from "Corticotropin-releasing factor increases Purkinje neuron excitability by modulating sodium, potassium, and Ih currents"

    Article Title: Corticotropin-releasing factor increases Purkinje neuron excitability by modulating sodium, potassium, and Ih currents

    Journal: Journal of Neurophysiology

    doi: 10.1152/jn.00745.2015

    CRF elicits voltage changes in PN. A : the average of 4 responses of PN to local application of CRF (8 μM, red bar) at four different holding potentials in the presence of tetrodotoxin (TTX). Note the reversal of the response between −40 mV and −50 mV. B : ZD-7288 blocks the TTX-insensitive depolarizing response. The integral of the depolarizing response measured from CRF onset over a duration of 5.5 s under control conditions ( n = 21), in the presence of TTX ( n = 13) and TTX + ZD-7288 ( n = 15) is shown. C , first trace: whole cell recording of the average response to CRF application in the presence of TTX. Second trace: 8 pulses of −40 pA for 0.5 s, delivered at 1 Hz during CRF response. The third trace is the subtraction of the first trace from the second trace. Red bar denote the CRF application. Note the reduction in the response to current injections during CRF application. Fourth trace is the current injection protocol. D : the average reduction in input resistance during CRF application in 5 PN. E : a representative example of seven superimposed traces of the hyperpolarizing response of PN to CRF applications obtained from different holding potentials (−75 to −35 mV) and aligned by the membrane voltage before the application. F : the result shown in E plotted as a function of the membrane potential (red curve) and similar curves measured from another 5 cells (black curves). 2 μM CRF was used in C – F .
    Figure Legend Snippet: CRF elicits voltage changes in PN. A : the average of 4 responses of PN to local application of CRF (8 μM, red bar) at four different holding potentials in the presence of tetrodotoxin (TTX). Note the reversal of the response between −40 mV and −50 mV. B : ZD-7288 blocks the TTX-insensitive depolarizing response. The integral of the depolarizing response measured from CRF onset over a duration of 5.5 s under control conditions ( n = 21), in the presence of TTX ( n = 13) and TTX + ZD-7288 ( n = 15) is shown. C , first trace: whole cell recording of the average response to CRF application in the presence of TTX. Second trace: 8 pulses of −40 pA for 0.5 s, delivered at 1 Hz during CRF response. The third trace is the subtraction of the first trace from the second trace. Red bar denote the CRF application. Note the reduction in the response to current injections during CRF application. Fourth trace is the current injection protocol. D : the average reduction in input resistance during CRF application in 5 PN. E : a representative example of seven superimposed traces of the hyperpolarizing response of PN to CRF applications obtained from different holding potentials (−75 to −35 mV) and aligned by the membrane voltage before the application. F : the result shown in E plotted as a function of the membrane potential (red curve) and similar curves measured from another 5 cells (black curves). 2 μM CRF was used in C – F .

    Techniques Used: Injection

    4) Product Images from "Regional and Developmental Differences in Na+ Currents in Vestibular Primary Afferent Neurons"

    Article Title: Regional and Developmental Differences in Na+ Currents in Vestibular Primary Afferent Neurons

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2018.00423

    A TTX-insensitive current is present in immature calyces. (A) Control I Na and currents remaining in the presence of 200 nM TTX at membrane potentials above –60 mV in a P8 calyx (note different current scales for left and right panels). Voltage protocol similar to Figure 2 : a 40 ms step to –130 mV from a holding potential of –80 mV was followed by a series of 40 ms depolarizing steps in 10 mV increments from –90 to +20 mV. (B) Control (black) and response to 200 nM TTX (red) for nine calyces (P5–11) are shown in the IV plot of peak inward currents for steps between –80 and 0 mV. (C) Left panel: the residual current following 200 nM TTX is blocked in 1 μM TTX and the block reverses with washout in a P6 calyx. Currents in response to a voltage step from –130 to –30 mV. Right panel: 1 μM JZTX-III blocks a component of I Na and the remaining current is abolished following application of 1 μM JZTX-III plus 200 nM TTX in a P7 calyx. Currents in response to a voltage step from –130 to –50 mV. (D) Summary for a group of 12 cells perfused with 200 nM TTX (six cells at P5–6, six at P8–11). A group of cells exposed to 1 μM TTX ( n = 8, one cell at P6, seven cells at P7–10) I Na was abolished in 1 μM JZTX-III and 200 nM TTX ( n = 4, two cells at P6 and two cells at P7). Peak inward current was measured at –50 mV step. I Na tended to be larger at younger ages as shown by distributions. In the presence of 200 nM TTX, I Na decreased from –3.9 (1.8) to –0.25 (0.5) nA ( P
    Figure Legend Snippet: A TTX-insensitive current is present in immature calyces. (A) Control I Na and currents remaining in the presence of 200 nM TTX at membrane potentials above –60 mV in a P8 calyx (note different current scales for left and right panels). Voltage protocol similar to Figure 2 : a 40 ms step to –130 mV from a holding potential of –80 mV was followed by a series of 40 ms depolarizing steps in 10 mV increments from –90 to +20 mV. (B) Control (black) and response to 200 nM TTX (red) for nine calyces (P5–11) are shown in the IV plot of peak inward currents for steps between –80 and 0 mV. (C) Left panel: the residual current following 200 nM TTX is blocked in 1 μM TTX and the block reverses with washout in a P6 calyx. Currents in response to a voltage step from –130 to –30 mV. Right panel: 1 μM JZTX-III blocks a component of I Na and the remaining current is abolished following application of 1 μM JZTX-III plus 200 nM TTX in a P7 calyx. Currents in response to a voltage step from –130 to –50 mV. (D) Summary for a group of 12 cells perfused with 200 nM TTX (six cells at P5–6, six at P8–11). A group of cells exposed to 1 μM TTX ( n = 8, one cell at P6, seven cells at P7–10) I Na was abolished in 1 μM JZTX-III and 200 nM TTX ( n = 4, two cells at P6 and two cells at P7). Peak inward current was measured at –50 mV step. I Na tended to be larger at younger ages as shown by distributions. In the presence of 200 nM TTX, I Na decreased from –3.9 (1.8) to –0.25 (0.5) nA ( P

    Techniques Used: Blocking Assay

    5) Product Images from "The floor-plate of His is a non-neuronal electrical conduction pathway"

    Article Title: The floor-plate of His is a non-neuronal electrical conduction pathway

    Journal: bioRxiv

    doi: 10.1101/2020.05.23.111955

    Floor plate biphasic action potentials are triggered by the activation of nicotinic acetylcholine receptors in response to acetylcholine released by motoneurons. a , Example of recurrent spontaneous floor plate action potentials blocked after addition of the nicotinic acetylcholine receptor (nAChR) antagonists: Mecamylamine (50 µM) and d-Tubocurarine (5 µM). b , Example of current-clamp recording showing floor plate action potential evoked by electrical stimulation in control condition (black trace) and after addition of the nAChR antagonists (red trace). c , Example of current-clamp recording showing floor plate action potential evoked in control condition (black trace) and after addition of antagonists against ionotropic receptor for GABA (Gabazine 3 µM) and glutamate (DL-APV 200 µM and CNQX 20 µM). d , Example of current-clamp recording showing that floor plate action potential can be evoked by local application of 30 µM acetylcholine (left trace), even in the presence of TTX (1 µM) and antagonists to AMPA/Kainate glutamate receptor (CNQX 10 µM), NMDA glutamate receptor (DL-APV 200 µM), GABA A receptor (Gabazine 3 µM) and glycine receptor (strychnine 1 µM) (center trace). Floor plate action potential evoked by acetylcholine were blocked by the addition of nAChR antagonists (right trace). Note that the addition of TTX inhibited the fast component of the biphasic action potential (see Extended Data Figure 2 ). e , Confocal image of a coronal section from a ChAT:ChR2-YFP mouse embryo at E12.5 showing the expression of Channelrhodopsin2-YFP fusion protein (in green) in cholinergic motoneurons located in the ventro-dorsal horns and labelled with the vesicular acetylcholine transporter VAChT (in red). All cell nuclei were labelled using DAPI (in blue). f , Example of current-clamp recording from a ChAT:ChR2-YFP + motoneuron showing an action potential triggered by the opening of Channelrhodopsin 2 in response to blue light stimulation (470 nm). g , Example of current-clamp recording from a floor plate cell recorded in a ChAT:ChR2-YFP + fetal spinal cord showing how blue light stimulation could evoke a slow cholinergic depolarization and trigger a biphasic action potential that were blocked by the addition of nAChR antagonists.
    Figure Legend Snippet: Floor plate biphasic action potentials are triggered by the activation of nicotinic acetylcholine receptors in response to acetylcholine released by motoneurons. a , Example of recurrent spontaneous floor plate action potentials blocked after addition of the nicotinic acetylcholine receptor (nAChR) antagonists: Mecamylamine (50 µM) and d-Tubocurarine (5 µM). b , Example of current-clamp recording showing floor plate action potential evoked by electrical stimulation in control condition (black trace) and after addition of the nAChR antagonists (red trace). c , Example of current-clamp recording showing floor plate action potential evoked in control condition (black trace) and after addition of antagonists against ionotropic receptor for GABA (Gabazine 3 µM) and glutamate (DL-APV 200 µM and CNQX 20 µM). d , Example of current-clamp recording showing that floor plate action potential can be evoked by local application of 30 µM acetylcholine (left trace), even in the presence of TTX (1 µM) and antagonists to AMPA/Kainate glutamate receptor (CNQX 10 µM), NMDA glutamate receptor (DL-APV 200 µM), GABA A receptor (Gabazine 3 µM) and glycine receptor (strychnine 1 µM) (center trace). Floor plate action potential evoked by acetylcholine were blocked by the addition of nAChR antagonists (right trace). Note that the addition of TTX inhibited the fast component of the biphasic action potential (see Extended Data Figure 2 ). e , Confocal image of a coronal section from a ChAT:ChR2-YFP mouse embryo at E12.5 showing the expression of Channelrhodopsin2-YFP fusion protein (in green) in cholinergic motoneurons located in the ventro-dorsal horns and labelled with the vesicular acetylcholine transporter VAChT (in red). All cell nuclei were labelled using DAPI (in blue). f , Example of current-clamp recording from a ChAT:ChR2-YFP + motoneuron showing an action potential triggered by the opening of Channelrhodopsin 2 in response to blue light stimulation (470 nm). g , Example of current-clamp recording from a floor plate cell recorded in a ChAT:ChR2-YFP + fetal spinal cord showing how blue light stimulation could evoke a slow cholinergic depolarization and trigger a biphasic action potential that were blocked by the addition of nAChR antagonists.

    Techniques Used: Activation Assay, Expressing

    6) Product Images from "Impaired neural pathway in gastric muscles of patients with diabetes"

    Article Title: Impaired neural pathway in gastric muscles of patients with diabetes

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-24147-y

    Electrical field stimulation (EFS)-induced response of distal gastric circular muscle strips after serial administration of atropine, MRS2500, N-nitro-L-arginine (L-NNA), and tetrodotoxin (TTX). ( a and b ) Atropine decreased the peak and TTX decreased the peak further in both the control subjects and diabetic patients. ( c ) MRS2500 increased the nadir but L-NNA abolished relaxation in the control subjects. ( d ) Also in the diabetic patients, L-NNA abolished relaxation. The Wilcoxon signed-rank test was used to evaluate the effects of each drug by compare values to the previous one.
    Figure Legend Snippet: Electrical field stimulation (EFS)-induced response of distal gastric circular muscle strips after serial administration of atropine, MRS2500, N-nitro-L-arginine (L-NNA), and tetrodotoxin (TTX). ( a and b ) Atropine decreased the peak and TTX decreased the peak further in both the control subjects and diabetic patients. ( c ) MRS2500 increased the nadir but L-NNA abolished relaxation in the control subjects. ( d ) Also in the diabetic patients, L-NNA abolished relaxation. The Wilcoxon signed-rank test was used to evaluate the effects of each drug by compare values to the previous one.

    Techniques Used:

    7) Product Images from "Selective Regulation of GluA Subunit Synthesis and AMPA Receptor-Mediated Synaptic Function and Plasticity by the Translation Repressor 4E-BP2 in Hippocampal Pyramidal Cells"

    Article Title: Selective Regulation of GluA Subunit Synthesis and AMPA Receptor-Mediated Synaptic Function and Plasticity by the Translation Repressor 4E-BP2 in Hippocampal Pyramidal Cells

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3264-12.2013

    Facilitation of AMPAR-mediated unitary excitatory synaptic transmission in 4E-BP2 −/− mice. A , Increased spontaneous miniature synaptic activity in 4E-BP2 −/− mice. Top: Continuous 1 s recordings (in TTX, AP-5, and gabazine), showing more frequent and larger amplitude mEPSCs in pyramidal cells of 4E-BP2 −/− mice. Bottom: Summary bar graphs for all cells, showing a greater mEPSC amplitude and frequency in slices from 4E-BP2 −/− mice. B , Representative EPSCs evoked by minimal stimulation in pyramidal neurons illustrating larger responses in slices from 4E-BP2 −/− mice. Left: Superimposed 20 successive traces (EPSC successes + failures; gray) and mean response of 100 events (including failures; black). Middle: Mean EPSC pairs evoked by paired-pulse stimulation (50 ms interpulse interval), showing similar paired-pulse facilitation in 4E-BP2 −/− mice. The first and second EPSCs are superimposed on the right. Right: Superimposed scaled EPSC pairs showing that paired-pulse ratio is unchanged in slices from 4E-BP2 −/− mice. C , Summary bar graphs showing facilitation of EPSC potency (mean EPSC without failures) and similar failure rate and paired-pulse ratio in 4E-BP2 −/− mice. Data are mean ± SEM. * p
    Figure Legend Snippet: Facilitation of AMPAR-mediated unitary excitatory synaptic transmission in 4E-BP2 −/− mice. A , Increased spontaneous miniature synaptic activity in 4E-BP2 −/− mice. Top: Continuous 1 s recordings (in TTX, AP-5, and gabazine), showing more frequent and larger amplitude mEPSCs in pyramidal cells of 4E-BP2 −/− mice. Bottom: Summary bar graphs for all cells, showing a greater mEPSC amplitude and frequency in slices from 4E-BP2 −/− mice. B , Representative EPSCs evoked by minimal stimulation in pyramidal neurons illustrating larger responses in slices from 4E-BP2 −/− mice. Left: Superimposed 20 successive traces (EPSC successes + failures; gray) and mean response of 100 events (including failures; black). Middle: Mean EPSC pairs evoked by paired-pulse stimulation (50 ms interpulse interval), showing similar paired-pulse facilitation in 4E-BP2 −/− mice. The first and second EPSCs are superimposed on the right. Right: Superimposed scaled EPSC pairs showing that paired-pulse ratio is unchanged in slices from 4E-BP2 −/− mice. C , Summary bar graphs showing facilitation of EPSC potency (mean EPSC without failures) and similar failure rate and paired-pulse ratio in 4E-BP2 −/− mice. Data are mean ± SEM. * p

    Techniques Used: Transmission Assay, Mouse Assay, Activity Assay, Mass Spectrometry

    8) Product Images from "Fxr1 regulates sleep and synaptic homeostasis"

    Article Title: Fxr1 regulates sleep and synaptic homeostasis

    Journal: The EMBO Journal

    doi: 10.15252/embj.2019103864

    Fxr1 protein expression is decreased during homeostatic synaptic upscaling Western blot analysis of Fxr1 during (A), TTX (48 h treatment) induced upscaling ( n = 6 in each condition) and (B), BIC (48 h treatment) induced downscaling ( n = 8 in each condition) of primary postnatal cortical cultures. Student's t ‐test * P
    Figure Legend Snippet: Fxr1 protein expression is decreased during homeostatic synaptic upscaling Western blot analysis of Fxr1 during (A), TTX (48 h treatment) induced upscaling ( n = 6 in each condition) and (B), BIC (48 h treatment) induced downscaling ( n = 8 in each condition) of primary postnatal cortical cultures. Student's t ‐test * P

    Techniques Used: Expressing, Western Blot

    Expression of GluA2 during synaptic scaling Quantification of the % of infection by (A) AAV1 Syn GFP ( n = 3) or (B) AAV1 Syn GFP‐Fxr1 ( n = 3). Surface expression of GluA2 during upscaling (Veh n = 28, TTX n = 29). Expression of total GluA2 during upscaling (Veh n = 7, TTX n = 6). Surface expression of GluA2 during downscaling (Veh n = 18, BIC n = 18). Expression of total GluA2 during downscaling (Veh n = 4, BIC n = 4). Data information: Error bars are SEM.
    Figure Legend Snippet: Expression of GluA2 during synaptic scaling Quantification of the % of infection by (A) AAV1 Syn GFP ( n = 3) or (B) AAV1 Syn GFP‐Fxr1 ( n = 3). Surface expression of GluA2 during upscaling (Veh n = 28, TTX n = 29). Expression of total GluA2 during upscaling (Veh n = 7, TTX n = 6). Surface expression of GluA2 during downscaling (Veh n = 18, BIC n = 18). Expression of total GluA2 during downscaling (Veh n = 4, BIC n = 4). Data information: Error bars are SEM.

    Techniques Used: Expressing, Infection

    The decrease in Fxr1 expression is necessary and sufficient for induction of multiplicative upscaling Cumulative probability plots of mEPSC amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical control neurons after 48 h of 1 μM TTX or Veh exposure (Ctrl/Veh n = 16 and Ctrl/TTX n = 17). A linear fit of Ctrl/TTX and Ctrl/Veh amplitudes. The degrees of overlap between Ctrl/TTX and Ctrl/Veh data were assessed using various scaling factors. The largest nonsignificant P ‐value was obtained with 1.47 scaling factor. Cumulative probability plots of the mEPSC amplitude of Ctrl/Veh, Ctrl/TTX, and Ctrl/TTX divided by scaling factor 1.47, which yielded the maximum overlap with Ctrl/Veh data. Cumulative probability plots of mEPSC amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical neurons after 48 h of 1 μM TTX or Veh exposure, (E) Fxr1 overexpressing neurons (Fxr1/Veh n = 10 and Fxr1/TTX n = 8), (F) Gsk3 KO neurons (Gsk3KO/Veh n = 10 and Gsk3KO/TTX n = 11), (G) Fxr1 KO neurons (Fxr1KO/Veh n = 16 and Fxr1KO/TTX n = 8). A linear fit of Fxr1 KO/Veh and Ctrl/Veh amplitudes. The degrees of overlap between Fxr1 KO/Veh and Ctrl/Veh data were assessed using various scaling factors. The largest nonsignificant P ‐value was obtained with 1.27 scaling factor. Cumulative probability plots of the mEPSC amplitude of Ctrl/Veh, Fxr1 KO/Veh, and Fxr1 KO/Veh divided by scaling factor 1.27, which yielded the maximum overlap with Ctrl/Veh data. Cumulative probability plots of mEPSC amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical Gsk3 and Fxr1 KO neurons after 48 h of 1 μM TTX or Veh exposure (Gsk3/Fxr1KO/Veh n = 8 and Gsk3/Fxr1KO/TTX n = 11). A linear fit of Gsk3/Fxr1 KO/Veh and Ctrl/Veh amplitudes. The degrees of overlap between Gsk3/Fxr1 KO/Veh and Ctrl/Veh data were assessed using various scaling factors. The largest nonsignificant P ‐value was obtained with 1.68 scaling factor. Cumulative probability plots of the mEPSC amplitude of Ctrl/Veh, Gsk3/Fxr1 KO/Veh, and Gsk3/Fxr1 KO/Veh divided by scaling factor 1.68, which yielded the maximum overlap with Ctrl/Veh data. mEPSC mean amplitude of cultured cortical neurons after 48 h of 1 μM TTX or Veh exposure. One‐way ANOVA with Bonferroni's multiple comparison test * P
    Figure Legend Snippet: The decrease in Fxr1 expression is necessary and sufficient for induction of multiplicative upscaling Cumulative probability plots of mEPSC amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical control neurons after 48 h of 1 μM TTX or Veh exposure (Ctrl/Veh n = 16 and Ctrl/TTX n = 17). A linear fit of Ctrl/TTX and Ctrl/Veh amplitudes. The degrees of overlap between Ctrl/TTX and Ctrl/Veh data were assessed using various scaling factors. The largest nonsignificant P ‐value was obtained with 1.47 scaling factor. Cumulative probability plots of the mEPSC amplitude of Ctrl/Veh, Ctrl/TTX, and Ctrl/TTX divided by scaling factor 1.47, which yielded the maximum overlap with Ctrl/Veh data. Cumulative probability plots of mEPSC amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical neurons after 48 h of 1 μM TTX or Veh exposure, (E) Fxr1 overexpressing neurons (Fxr1/Veh n = 10 and Fxr1/TTX n = 8), (F) Gsk3 KO neurons (Gsk3KO/Veh n = 10 and Gsk3KO/TTX n = 11), (G) Fxr1 KO neurons (Fxr1KO/Veh n = 16 and Fxr1KO/TTX n = 8). A linear fit of Fxr1 KO/Veh and Ctrl/Veh amplitudes. The degrees of overlap between Fxr1 KO/Veh and Ctrl/Veh data were assessed using various scaling factors. The largest nonsignificant P ‐value was obtained with 1.27 scaling factor. Cumulative probability plots of the mEPSC amplitude of Ctrl/Veh, Fxr1 KO/Veh, and Fxr1 KO/Veh divided by scaling factor 1.27, which yielded the maximum overlap with Ctrl/Veh data. Cumulative probability plots of mEPSC amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical Gsk3 and Fxr1 KO neurons after 48 h of 1 μM TTX or Veh exposure (Gsk3/Fxr1KO/Veh n = 8 and Gsk3/Fxr1KO/TTX n = 11). A linear fit of Gsk3/Fxr1 KO/Veh and Ctrl/Veh amplitudes. The degrees of overlap between Gsk3/Fxr1 KO/Veh and Ctrl/Veh data were assessed using various scaling factors. The largest nonsignificant P ‐value was obtained with 1.68 scaling factor. Cumulative probability plots of the mEPSC amplitude of Ctrl/Veh, Gsk3/Fxr1 KO/Veh, and Gsk3/Fxr1 KO/Veh divided by scaling factor 1.68, which yielded the maximum overlap with Ctrl/Veh data. mEPSC mean amplitude of cultured cortical neurons after 48 h of 1 μM TTX or Veh exposure. One‐way ANOVA with Bonferroni's multiple comparison test * P

    Techniques Used: Expressing, Cell Culture

    Fxr1 suppresses the increase in surface GluA1 during synaptic upscaling Schematic of high‐efficiency infection of neuronal cultures by AAV1 viruses followed by detection of AMPA receptor subunits. Western blot analysis of total GluA1 expression in Ctrl or Fxr1 overexpression (Fxr1) condition during upscaling (Ctrl/Veh n = 5, Ctrl/TTX n = 5, Fxr1/Veh n = 6, Fxr1/Veh n = 6). One‐way ANOVA with Dunnett's multiple comparison test *** P
    Figure Legend Snippet: Fxr1 suppresses the increase in surface GluA1 during synaptic upscaling Schematic of high‐efficiency infection of neuronal cultures by AAV1 viruses followed by detection of AMPA receptor subunits. Western blot analysis of total GluA1 expression in Ctrl or Fxr1 overexpression (Fxr1) condition during upscaling (Ctrl/Veh n = 5, Ctrl/TTX n = 5, Fxr1/Veh n = 6, Fxr1/Veh n = 6). One‐way ANOVA with Dunnett's multiple comparison test *** P

    Techniques Used: Infection, Western Blot, Expressing, Over Expression

    9) Product Images from "Extracellular α-synuclein levels are regulated by neuronal activity"

    Article Title: Extracellular α-synuclein levels are regulated by neuronal activity

    Journal: Molecular Neurodegeneration

    doi: 10.1186/s13024-018-0241-0

    Synaptic vesicle exocytosis is associated with α-synuclein release. a Schematic illustration of target molecules for the compounds used in the experiment. b α-Latrotoxin (αLTX, 0.5 nM) increased α-synuclein release, which was blocked by 4 mM EGTA. N = 8. c αLTX significantly increased α-synuclein release in the presence or absence of 1 μM TTX, 50 μM AP5 and 10 μM NBQX. N = 16–32. d LDH activities in the media were not altered by indicated pharmacological treatments. N = 16–24. e Cellular α-synuclein levels were not altered by indicate pharmacological treatments. N = 8. mean ± SEM, *** p
    Figure Legend Snippet: Synaptic vesicle exocytosis is associated with α-synuclein release. a Schematic illustration of target molecules for the compounds used in the experiment. b α-Latrotoxin (αLTX, 0.5 nM) increased α-synuclein release, which was blocked by 4 mM EGTA. N = 8. c αLTX significantly increased α-synuclein release in the presence or absence of 1 μM TTX, 50 μM AP5 and 10 μM NBQX. N = 16–32. d LDH activities in the media were not altered by indicated pharmacological treatments. N = 16–24. e Cellular α-synuclein levels were not altered by indicate pharmacological treatments. N = 8. mean ± SEM, *** p

    Techniques Used:

    10) Product Images from "Dynamics of GnRH Neuron Ionotropic GABA and Glutamate Synaptic Receptors Are Unchanged during Estrogen Positive and Negative Feedback in Female Mice"

    Article Title: Dynamics of GnRH Neuron Ionotropic GABA and Glutamate Synaptic Receptors Are Unchanged during Estrogen Positive and Negative Feedback in Female Mice

    Journal: eNeuro

    doi: 10.1523/ENEURO.0259-17.2017

    Miniature GABA A receptor PSCs (mGPSCs) recorded from GnRH neurons do not change in the different mouse models of estrogen negative and positive feedback. A - F , Representative examples of mGPSC current recordings in the presence of AP5 (50 μM), CNQX (10 μM), and TTX (0.5 μM) taken from diestrous, OVX, OVX + E, and OVX + E+E mice. Underneath each trace an enlarged time scale of one mGPSC (*) is shown. C , F , mGPSCs are abolished by GABAzine (5 μM). G , H , Cumulative plots of the average inter-mGPSC interval ( G ) and amplitude ( H ) in the different experimental groups (for clarity only positive SEM is shown). N = 7 for each group. Cell numbers are shown in parenthesis. Numbers of PSCs analyzed is given in Table 1 . No statistically significant differences were detected for any parameter between groups.
    Figure Legend Snippet: Miniature GABA A receptor PSCs (mGPSCs) recorded from GnRH neurons do not change in the different mouse models of estrogen negative and positive feedback. A - F , Representative examples of mGPSC current recordings in the presence of AP5 (50 μM), CNQX (10 μM), and TTX (0.5 μM) taken from diestrous, OVX, OVX + E, and OVX + E+E mice. Underneath each trace an enlarged time scale of one mGPSC (*) is shown. C , F , mGPSCs are abolished by GABAzine (5 μM). G , H , Cumulative plots of the average inter-mGPSC interval ( G ) and amplitude ( H ) in the different experimental groups (for clarity only positive SEM is shown). N = 7 for each group. Cell numbers are shown in parenthesis. Numbers of PSCs analyzed is given in Table 1 . No statistically significant differences were detected for any parameter between groups.

    Techniques Used: Mouse Assay

    Miniature excitatory glutamate receptor PSCs (mEPSCs) recorded from GnRH neurons do not change in the different mouse models of estrogen negative and positive feedback. A - F , Representative examples of mEPSCs current recordings in the presence of GABAzine (5 μM) and TTX (0.5 μM) taken from diestrous, OVX, OVX + E, and OVX + E+E mice. Underneath each trace an enlarged time scale of one mEPSC (*) is shown. C , F , mEPSCs are abolished by AP5 (50 μM) + CNQX (10 μM). G , H , Cumulative plots of the average inter-mEPSC interval ( G ) and amplitude ( H ) in the different experimental groups (for clarity only positive SEM is shown). N = 5 for each group. Cell numbers are shown in parenthesis. Numbers of PSCs analyzed is given in Table 2 . No statistically significant differences were detected for any parameter between groups.
    Figure Legend Snippet: Miniature excitatory glutamate receptor PSCs (mEPSCs) recorded from GnRH neurons do not change in the different mouse models of estrogen negative and positive feedback. A - F , Representative examples of mEPSCs current recordings in the presence of GABAzine (5 μM) and TTX (0.5 μM) taken from diestrous, OVX, OVX + E, and OVX + E+E mice. Underneath each trace an enlarged time scale of one mEPSC (*) is shown. C , F , mEPSCs are abolished by AP5 (50 μM) + CNQX (10 μM). G , H , Cumulative plots of the average inter-mEPSC interval ( G ) and amplitude ( H ) in the different experimental groups (for clarity only positive SEM is shown). N = 5 for each group. Cell numbers are shown in parenthesis. Numbers of PSCs analyzed is given in Table 2 . No statistically significant differences were detected for any parameter between groups.

    Techniques Used: Mouse Assay

    11) Product Images from "Fxr1 regulates sleep and synaptic homeostasis"

    Article Title: Fxr1 regulates sleep and synaptic homeostasis

    Journal: bioRxiv

    doi: 10.1101/709345

    The decrease in Fxr1 expression is necessary and sufficient for induction of multiplicative upscaling. a , Cumulative probability plots of mEPSCs amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical control neurons after 48 hours of 1 µM TTX or Veh exposure (Ctrl/Veh n=16 and Ctrl/TTX n=17). b , A linear fit of Ctrl/TTX and Ctrl/Veh amplitudes. c , The degrees of overlap between Ctrl/TTX and Ctrl/Veh data were assessed using various scaling factors. The largest nonsignificant p-value was obtained with 1.47 scaling factor. d , Cumulative probability plots of the mEPSCs amplitude of Ctrl/Veh, Ctrl/TTX and Ctrl/TTX divided by scaling factor 1.47, which yielded the maximum overlap with Ctrl/Veh data. e-g , Cumulative probability plots of mEPSCs amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical neurons after 48 hours of 1 µM TTX or Veh exposure e , Fxr1P overexpressing neurons (Fxr1P over/Veh n=10 and Fxr1P over/TTX n=8), f , Gsk3 KO neurons (Gsk3KO/Veh n=10 and Gsk3KO/TTX n=11), g , Fxr1 KO neurons (Fxr1KO/Veh n=16 and Fxr1KO/TTX n=8). h , A linear fit of Fxr1 KO/Veh and Ctrl/Veh amplitudes. i , The degrees of overlap between Fxr1 KO/Veh and Ctrl/Veh data were assessed using various scaling factors. The largest non-significant p-value was obtained with 1.27 scaling factor. j , Cumulative probability plots of the mEPSCs amplitude of Ctrl/Veh, Fxr1 KO/Veh and Fxr1 KO/Veh divided by scaling factor 1.27, which yielded the maximum overlap with Ctrl/Veh data. k , Cumulative probability plots of mEPSCs amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical Gsk3 and Fxr1 KO neurons after 48 hours of 1 µM TTX or Veh exposure (Gsk3/Fxr1KO/Veh n=8 and Gsk3/Fxr1KO/TTX n=11). l , A linear fit of Gsk3/Fxr1 KO/Veh and Ctrl/Veh amplitudes. m , The degrees of overlap between Gsk3/Fxr1 KO/Veh and Ctrl/Veh data were assessed using various scaling factors. The largest non-significant p-value was obtained with 1.68 scaling factor. n , Cumulative probability plots of the mEPSCs amplitude of Ctrl/Veh, Gsk3/Fxr1 KO/Veh and Gsk3/Fxr1 KO/Veh divided by scaling factor 1.68, which yielded the maximum overlap with Ctrl/Veh data. o , mEPSC amplitude of cultured cortical neurons after 48 hours of 1 µM TTX or Veh exposure. One way Anova with Bonferroni’s Multiple Comparison Test *p
    Figure Legend Snippet: The decrease in Fxr1 expression is necessary and sufficient for induction of multiplicative upscaling. a , Cumulative probability plots of mEPSCs amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical control neurons after 48 hours of 1 µM TTX or Veh exposure (Ctrl/Veh n=16 and Ctrl/TTX n=17). b , A linear fit of Ctrl/TTX and Ctrl/Veh amplitudes. c , The degrees of overlap between Ctrl/TTX and Ctrl/Veh data were assessed using various scaling factors. The largest nonsignificant p-value was obtained with 1.47 scaling factor. d , Cumulative probability plots of the mEPSCs amplitude of Ctrl/Veh, Ctrl/TTX and Ctrl/TTX divided by scaling factor 1.47, which yielded the maximum overlap with Ctrl/Veh data. e-g , Cumulative probability plots of mEPSCs amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical neurons after 48 hours of 1 µM TTX or Veh exposure e , Fxr1P overexpressing neurons (Fxr1P over/Veh n=10 and Fxr1P over/TTX n=8), f , Gsk3 KO neurons (Gsk3KO/Veh n=10 and Gsk3KO/TTX n=11), g , Fxr1 KO neurons (Fxr1KO/Veh n=16 and Fxr1KO/TTX n=8). h , A linear fit of Fxr1 KO/Veh and Ctrl/Veh amplitudes. i , The degrees of overlap between Fxr1 KO/Veh and Ctrl/Veh data were assessed using various scaling factors. The largest non-significant p-value was obtained with 1.27 scaling factor. j , Cumulative probability plots of the mEPSCs amplitude of Ctrl/Veh, Fxr1 KO/Veh and Fxr1 KO/Veh divided by scaling factor 1.27, which yielded the maximum overlap with Ctrl/Veh data. k , Cumulative probability plots of mEPSCs amplitude (500 events per cell) and representative examples of mEPSCs (left panel) recorded from cultured cortical Gsk3 and Fxr1 KO neurons after 48 hours of 1 µM TTX or Veh exposure (Gsk3/Fxr1KO/Veh n=8 and Gsk3/Fxr1KO/TTX n=11). l , A linear fit of Gsk3/Fxr1 KO/Veh and Ctrl/Veh amplitudes. m , The degrees of overlap between Gsk3/Fxr1 KO/Veh and Ctrl/Veh data were assessed using various scaling factors. The largest non-significant p-value was obtained with 1.68 scaling factor. n , Cumulative probability plots of the mEPSCs amplitude of Ctrl/Veh, Gsk3/Fxr1 KO/Veh and Gsk3/Fxr1 KO/Veh divided by scaling factor 1.68, which yielded the maximum overlap with Ctrl/Veh data. o , mEPSC amplitude of cultured cortical neurons after 48 hours of 1 µM TTX or Veh exposure. One way Anova with Bonferroni’s Multiple Comparison Test *p

    Techniques Used: Expressing, Cell Culture

    Fxr1 suppresses the increase of surface GluA1 during synaptic upscaling a , Schematic of high-efficiency infection of neuronal cultures by AAV1 viruses followed by detection of AMPA receptor subunits. b , Western blot analysis of total GluA1 expression in Ctrl or Fxr1over condition during upscaling (Ctrl/Veh n=5, Ctrl/TTX n=5, Fxr1over/Veh n=6, Fxr1over/Veh n=6). One way Anova with Dunnett’s Multiple Comparison Test ***p
    Figure Legend Snippet: Fxr1 suppresses the increase of surface GluA1 during synaptic upscaling a , Schematic of high-efficiency infection of neuronal cultures by AAV1 viruses followed by detection of AMPA receptor subunits. b , Western blot analysis of total GluA1 expression in Ctrl or Fxr1over condition during upscaling (Ctrl/Veh n=5, Ctrl/TTX n=5, Fxr1over/Veh n=6, Fxr1over/Veh n=6). One way Anova with Dunnett’s Multiple Comparison Test ***p

    Techniques Used: Infection, Western Blot, Expressing

    Fxr1 protein expression is decreased during homeostatic synaptic upscaling. Western blot analysis of Fxr1 during a , TTX induced upscaling (n=6 in each condition) and b , BIC induced downscaling (n=4 in each condition) of primary postnatal cortical cultures. Student’s T-test *p
    Figure Legend Snippet: Fxr1 protein expression is decreased during homeostatic synaptic upscaling. Western blot analysis of Fxr1 during a , TTX induced upscaling (n=6 in each condition) and b , BIC induced downscaling (n=4 in each condition) of primary postnatal cortical cultures. Student’s T-test *p

    Techniques Used: Expressing, Western Blot

    12) Product Images from "Barbaloin inhibits ventricular arrhythmias in rabbits by modulating voltage-gated ion channels"

    Article Title: Barbaloin inhibits ventricular arrhythmias in rabbits by modulating voltage-gated ion channels

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2017.93

    Effects of barbaloin on ATX II-induced late sodium current ( I Na.L ) enhancements and under normal conditions in rabbit ventricular myocytes. (A) Representative single traces of the effects of 4 μmol/L TTX on ATX II-induced I Na.L enhancement. (B) Representative single traces showing that barbaloin exerted reversible inhibitory effects on ATX II-induced I Na.L enhancement. (C) Representative single traces of the effects of 200 μmol/L barbaloin on I Na.L under normal conditions. (D) Single traces of the effects of barbaloin on ATX II-induced I Na.L enhancements at a membrane potential of −20 mV. (E) Representative traces of the effects of barbaloin on ATX II-induced I Na.L enhancements at membrane potentials of −80, −60, −50, −40 and −20 mV. (F) Bar graphs showing the mean ATX II-increased I Na.L percentage values for the control, 100 μmol/L barbaloin-treated and wash-out groups ( n =10. ** P
    Figure Legend Snippet: Effects of barbaloin on ATX II-induced late sodium current ( I Na.L ) enhancements and under normal conditions in rabbit ventricular myocytes. (A) Representative single traces of the effects of 4 μmol/L TTX on ATX II-induced I Na.L enhancement. (B) Representative single traces showing that barbaloin exerted reversible inhibitory effects on ATX II-induced I Na.L enhancement. (C) Representative single traces of the effects of 200 μmol/L barbaloin on I Na.L under normal conditions. (D) Single traces of the effects of barbaloin on ATX II-induced I Na.L enhancements at a membrane potential of −20 mV. (E) Representative traces of the effects of barbaloin on ATX II-induced I Na.L enhancements at membrane potentials of −80, −60, −50, −40 and −20 mV. (F) Bar graphs showing the mean ATX II-increased I Na.L percentage values for the control, 100 μmol/L barbaloin-treated and wash-out groups ( n =10. ** P

    Techniques Used:

    13) Product Images from "Enhanced prefrontal serotonin 5-HT1A currents in a mouse model of Williams-Beuren syndrome with low innate anxiety"

    Article Title: Enhanced prefrontal serotonin 5-HT1A currents in a mouse model of Williams-Beuren syndrome with low innate anxiety

    Journal: Journal of neurodevelopmental disorders

    doi: 10.1007/s11689-010-9044-5

    Serotonergic currents in layer V pyramidal neurons mediated by 5-HT 1A receptors in wildtype (WT) and Gtf2ird1 −/− mice. In neurons from each genotype, (1.) the 5-HT-elicited outward currents were suppressed by (2.) the selective antagonist of 5-HT 1A receptors, WAY-100635 (30 nM, 10 min). Gray bar indicates 5-HT application. All the above traces were recorded after application of TTX (2 µM, 10 min), indicating that the 5-HT outward currents were mediated directly by 5-HT 1A receptors located on the recorded cells
    Figure Legend Snippet: Serotonergic currents in layer V pyramidal neurons mediated by 5-HT 1A receptors in wildtype (WT) and Gtf2ird1 −/− mice. In neurons from each genotype, (1.) the 5-HT-elicited outward currents were suppressed by (2.) the selective antagonist of 5-HT 1A receptors, WAY-100635 (30 nM, 10 min). Gray bar indicates 5-HT application. All the above traces were recorded after application of TTX (2 µM, 10 min), indicating that the 5-HT outward currents were mediated directly by 5-HT 1A receptors located on the recorded cells

    Techniques Used: Mouse Assay

    14) Product Images from "Subunit- and pathway-specific localization of NMDA receptors and scaffolding proteins at ganglion cell synapses in rat retina"

    Article Title: Subunit- and pathway-specific localization of NMDA receptors and scaffolding proteins at ganglion cell synapses in rat retina

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    doi: 10.1523/JNEUROSCI.5602-08.2009

    Distinct NMDAR subtype contributions to sEPSCs at ON and OFF synapses. A, Average sEPSCs recorded form a morphologically identified ON RGC (V hold = −80 mV, 1 μM TTX, 100 μM D-serine, 0 [Mg 2+ ] o , inhibition blocked). Reducing glutamate uptake with the transporter antagonist TBOA (10 μM, green, 95 events averaged) conferred a slow component onto the sEPSC waveform compared to control (black, 135 events). This TBOA-induced component was reduced by the NR2B NMDAR-specific antagonist Ro-25,6981 (Ro, 1 μM, red, 106 events) and abolished completely by the pan-NMDAR antagonist CPP (10 μM, blue, 110 events). B , Summarized effects of TBOA, Ro-25,6981 and CPP in 5 ON RGCs. C , Average sEPSCs recorded from an identified OFF RGC in the same control conditions as above (black, 425 events). In the absence of TBOA, Ro-25,6981 (red, 202 events) had no effect on the sEPSC waveform but CPP blocked a slow component (blue, 88 events), indicating the presence of synaptic NMDARs that lack NR2B subunits. D , Summarized effects of Ro-25,6981 and CPP in 5 OFF RGCs. E , Average sEPSCs recorded from an identified OFF RGC in the same control conditions as above (black, 133 events). Addition of TBOA enhanced the sEPSC waveform (green, 110 events), revealing a component that was eliminated by Ro-25,6981 (red, 120 events). CPP reduced the sEPSC waveform further (blue, 100 events). F , Summarized effects of TBOA, Ro-25,6981 and CPP in 5 OFF RGCs. G , Average sEPSCs recorded from an identified OFF RGC in the presence of TBOA (with inhibition blocked), in control (2.5 mM) [Ca 2+ ] o (black, 89 events) or low (0.5 mM) [Ca 2+ ] o (gray, 59 events). H , Effects of changing [Ca 2+ ] o on sEPSC frequency and charge transfer in 4 OFF RGCs.
    Figure Legend Snippet: Distinct NMDAR subtype contributions to sEPSCs at ON and OFF synapses. A, Average sEPSCs recorded form a morphologically identified ON RGC (V hold = −80 mV, 1 μM TTX, 100 μM D-serine, 0 [Mg 2+ ] o , inhibition blocked). Reducing glutamate uptake with the transporter antagonist TBOA (10 μM, green, 95 events averaged) conferred a slow component onto the sEPSC waveform compared to control (black, 135 events). This TBOA-induced component was reduced by the NR2B NMDAR-specific antagonist Ro-25,6981 (Ro, 1 μM, red, 106 events) and abolished completely by the pan-NMDAR antagonist CPP (10 μM, blue, 110 events). B , Summarized effects of TBOA, Ro-25,6981 and CPP in 5 ON RGCs. C , Average sEPSCs recorded from an identified OFF RGC in the same control conditions as above (black, 425 events). In the absence of TBOA, Ro-25,6981 (red, 202 events) had no effect on the sEPSC waveform but CPP blocked a slow component (blue, 88 events), indicating the presence of synaptic NMDARs that lack NR2B subunits. D , Summarized effects of Ro-25,6981 and CPP in 5 OFF RGCs. E , Average sEPSCs recorded from an identified OFF RGC in the same control conditions as above (black, 133 events). Addition of TBOA enhanced the sEPSC waveform (green, 110 events), revealing a component that was eliminated by Ro-25,6981 (red, 120 events). CPP reduced the sEPSC waveform further (blue, 100 events). F , Summarized effects of TBOA, Ro-25,6981 and CPP in 5 OFF RGCs. G , Average sEPSCs recorded from an identified OFF RGC in the presence of TBOA (with inhibition blocked), in control (2.5 mM) [Ca 2+ ] o (black, 89 events) or low (0.5 mM) [Ca 2+ ] o (gray, 59 events). H , Effects of changing [Ca 2+ ] o on sEPSC frequency and charge transfer in 4 OFF RGCs.

    Techniques Used: Inhibition, Conditioned Place Preference

    15) Product Images from "Mechanistic insights into the detection of free fatty and bile acids by ileal glucagon-like peptide-1 secreting cells"

    Article Title: Mechanistic insights into the detection of free fatty and bile acids by ileal glucagon-like peptide-1 secreting cells

    Journal: Molecular Metabolism

    doi: 10.1016/j.molmet.2017.11.005

    Electrophysiological characterization of organoid-derived ileal L-cells. (A) Perforated-patch current clamp recording of an organoid-derived ileal L-cell, firing action potentials in response to depolarizing current injections. Current was injected to maintain the cell at −70 mV, and a series of 10 ms current pulses were applied, increasing in magnitude by 2 pA. The pulse protocol is illustrated below. (B) Perforated-patch current clamp recording of spontaneous action potential firing from an ileal L-cell. (C) Representative traces using the same protocol as in (A), before (Ci) and during application of 0.3 μM tetrodotoxin (TTX, Cii) and during application of TTX + 0.5 μM ω-agatoxin-IVA (Ciii). Dashed line represents the threshold of action potential firing. The insets show spontaneous action potential firing under the same treatment conditions. (Civ) Threshold for action potential firing (n = 5) and (Cv) % inhibition of action potential peak following application of channel blockers, expressed as a % of total block by application of TTX (0.3 μM) + Cd 2+ (100 μM). (Di) Inward current from a perforated-patch voltage clamp recording and sample traces following application of 0.3 μM TTX (Dii, orange trace) or 100 μM Cd 2+ (Diii, gray trace). Currents were elicited from a series of 70 ms voltage steps from −110 to +60 mV, from a holding potential of −80 mV. Only the current response to the +10 mV voltage step is shown and is illustrated below the current traces. (Div) Peak current amplitude of the fast and slow current components. Gene expression data of Scn (Ei) or Cacna (Eii) genes by RNA sequencing of FACS-sorted L-cells from mouse ileum (white circles) and colon (black circles). Individual data points represent fragments per kilobase of transcript per million mapped reads (FPKM) from 1 mouse. Mean values (n = 3) are presented as lines. (Fi) Superimposed Ca 2+ currents from an ileal L-cell before and during exposure to Ca 2+ channel blockers. Red trace represents calcium current ( I Ca ) recorded in the presence of ω-agatoxin-IVA (0.5 μM), pink trace represents I Ca recorded following subsequent application of isradipine (10 μM), and gray trace represents I Ca recorded following application of cadmium (Cd 2+ , 100 μM). Currents were elicited using the protocol described in (D) and only the current responses to the +10 mV voltage step are shown. (Fii) Contribution of Ca 2+ channel subtype to total Ca 2+ current measured. (Fiii) The peak I Ca –voltage relationship for a representative organoid ileal L-cell following application of Ca 2+ channel blockers. Statistical analysis performed using either by Wilcoxon matched-pairs signed rank test (Civ), one-way ANOVA with Tukey's multiple comparison (Cv) or multiple t-tests with Holm-Sidak multiple comparisons correction (E), p
    Figure Legend Snippet: Electrophysiological characterization of organoid-derived ileal L-cells. (A) Perforated-patch current clamp recording of an organoid-derived ileal L-cell, firing action potentials in response to depolarizing current injections. Current was injected to maintain the cell at −70 mV, and a series of 10 ms current pulses were applied, increasing in magnitude by 2 pA. The pulse protocol is illustrated below. (B) Perforated-patch current clamp recording of spontaneous action potential firing from an ileal L-cell. (C) Representative traces using the same protocol as in (A), before (Ci) and during application of 0.3 μM tetrodotoxin (TTX, Cii) and during application of TTX + 0.5 μM ω-agatoxin-IVA (Ciii). Dashed line represents the threshold of action potential firing. The insets show spontaneous action potential firing under the same treatment conditions. (Civ) Threshold for action potential firing (n = 5) and (Cv) % inhibition of action potential peak following application of channel blockers, expressed as a % of total block by application of TTX (0.3 μM) + Cd 2+ (100 μM). (Di) Inward current from a perforated-patch voltage clamp recording and sample traces following application of 0.3 μM TTX (Dii, orange trace) or 100 μM Cd 2+ (Diii, gray trace). Currents were elicited from a series of 70 ms voltage steps from −110 to +60 mV, from a holding potential of −80 mV. Only the current response to the +10 mV voltage step is shown and is illustrated below the current traces. (Div) Peak current amplitude of the fast and slow current components. Gene expression data of Scn (Ei) or Cacna (Eii) genes by RNA sequencing of FACS-sorted L-cells from mouse ileum (white circles) and colon (black circles). Individual data points represent fragments per kilobase of transcript per million mapped reads (FPKM) from 1 mouse. Mean values (n = 3) are presented as lines. (Fi) Superimposed Ca 2+ currents from an ileal L-cell before and during exposure to Ca 2+ channel blockers. Red trace represents calcium current ( I Ca ) recorded in the presence of ω-agatoxin-IVA (0.5 μM), pink trace represents I Ca recorded following subsequent application of isradipine (10 μM), and gray trace represents I Ca recorded following application of cadmium (Cd 2+ , 100 μM). Currents were elicited using the protocol described in (D) and only the current responses to the +10 mV voltage step are shown. (Fii) Contribution of Ca 2+ channel subtype to total Ca 2+ current measured. (Fiii) The peak I Ca –voltage relationship for a representative organoid ileal L-cell following application of Ca 2+ channel blockers. Statistical analysis performed using either by Wilcoxon matched-pairs signed rank test (Civ), one-way ANOVA with Tukey's multiple comparison (Cv) or multiple t-tests with Holm-Sidak multiple comparisons correction (E), p

    Techniques Used: Derivative Assay, Injection, Mass Spectrometry, Inhibition, Blocking Assay, Expressing, RNA Sequencing Assay, FACS

    16) Product Images from "Translocation of CaMKII to dendritic microtubules supports the plasticity of local synapses"

    Article Title: Translocation of CaMKII to dendritic microtubules supports the plasticity of local synapses

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201202058

    Activity-dependent translocation of αCaMKII to dendritic sites near synapses. (A) Hippocampal neuron (12 DIV) expressing mGFP-αCaMKII and mCherry imaged before, during, and after stimulation with 0Mg 2+ /Gly for 5 min. (B, i) Pseudocolor image from dendrite in A. Kymograph (ii–iii) and time-lapse (iv) analyses of the change in the fluorescent intensity ratio (mGFP-αCaMKII/mCherry) over time across the dendrite (i–ii, black line) and across the spine (iii, yellow box, red line). Stimulation period is indicated (Stim). (iv) n = 4 spines and subdendritic regions from the neuron shown in A. Red and white arrows (or brackets) point, respectively, to synaptic and dendritic sites where CaMKII translocated. Bars: (neuron) 10 µm; (dendrite) 5 µm. (C) Normalized ratio (±SEM) of dendritic segment where mGFP-αCaMKII accumulated over total dendritic length after a 5-min 0Mg 2+ /Gly stimulation (Vehicle), in the presence of 50 µM AP5, 1 μM TTX, 10 μM Cd 2+ , or 10 μM CPA. n = 7–24 neurons per condition. *, P
    Figure Legend Snippet: Activity-dependent translocation of αCaMKII to dendritic sites near synapses. (A) Hippocampal neuron (12 DIV) expressing mGFP-αCaMKII and mCherry imaged before, during, and after stimulation with 0Mg 2+ /Gly for 5 min. (B, i) Pseudocolor image from dendrite in A. Kymograph (ii–iii) and time-lapse (iv) analyses of the change in the fluorescent intensity ratio (mGFP-αCaMKII/mCherry) over time across the dendrite (i–ii, black line) and across the spine (iii, yellow box, red line). Stimulation period is indicated (Stim). (iv) n = 4 spines and subdendritic regions from the neuron shown in A. Red and white arrows (or brackets) point, respectively, to synaptic and dendritic sites where CaMKII translocated. Bars: (neuron) 10 µm; (dendrite) 5 µm. (C) Normalized ratio (±SEM) of dendritic segment where mGFP-αCaMKII accumulated over total dendritic length after a 5-min 0Mg 2+ /Gly stimulation (Vehicle), in the presence of 50 µM AP5, 1 μM TTX, 10 μM Cd 2+ , or 10 μM CPA. n = 7–24 neurons per condition. *, P

    Techniques Used: Activity Assay, Translocation Assay, Expressing

    17) Product Images from "Functional segregation of voltage-activated calcium channels in motoneurons of the dorsal motor nucleus of the vagus"

    Article Title: Functional segregation of voltage-activated calcium channels in motoneurons of the dorsal motor nucleus of the vagus

    Journal: Journal of Neurophysiology

    doi: 10.1152/jn.00432.2014

    A-type Kv4 K + currents slow pacemaking of DMV neurons. A , left : example of measurements of the activation (black) and inactivation (gray) of the transient K + current in the presence of TTX. A , right : normalized Boltzman sigmoidal functions describing the voltage dependence of activation and inactivation of the Kv4 current, and the distribution of the voltages of half activation ( n = 13) and inactivation ( n = 15), reveal a window Kv4 current that spans the typical subthreshold range of the voltage trajectory of DMV neurons during pacemaking. B , left : Time course of the reduction in the peak A current as a function of time after membrane rupture in whole cell configuration. B , right : comparison of the peak A current at membrane rupture vs. past 4 min from rupture. C , left : time course of the instantaneous firing rate of a DMV neuron in cell-attached mode before membrane rupture and after whole cell configuration was achieved. B , right : comparison of autonomous discharge rate before and after rupturing the membrane. D : brief application of nominally 1–2 μM phrixotoxin-2 (PaTX-2), a selective Kv4 antagonist, transiently increases the firing rate of DMV neurons recorded in the loose-patch configuration (shown on a logarithmic scale). * P
    Figure Legend Snippet: A-type Kv4 K + currents slow pacemaking of DMV neurons. A , left : example of measurements of the activation (black) and inactivation (gray) of the transient K + current in the presence of TTX. A , right : normalized Boltzman sigmoidal functions describing the voltage dependence of activation and inactivation of the Kv4 current, and the distribution of the voltages of half activation ( n = 13) and inactivation ( n = 15), reveal a window Kv4 current that spans the typical subthreshold range of the voltage trajectory of DMV neurons during pacemaking. B , left : Time course of the reduction in the peak A current as a function of time after membrane rupture in whole cell configuration. B , right : comparison of the peak A current at membrane rupture vs. past 4 min from rupture. C , left : time course of the instantaneous firing rate of a DMV neuron in cell-attached mode before membrane rupture and after whole cell configuration was achieved. B , right : comparison of autonomous discharge rate before and after rupturing the membrane. D : brief application of nominally 1–2 μM phrixotoxin-2 (PaTX-2), a selective Kv4 antagonist, transiently increases the firing rate of DMV neurons recorded in the loose-patch configuration (shown on a logarithmic scale). * P

    Techniques Used: Activation Assay

    18) Product Images from "Impaired neural pathway in gastric muscles of patients with diabetes"

    Article Title: Impaired neural pathway in gastric muscles of patients with diabetes

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-24147-y

    Electrical field stimulation (EFS)-induced response of distal gastric circular muscle strips after serial administration of atropine, MRS2500, N-nitro-L-arginine (L-NNA), and tetrodotoxin (TTX). ( a and b ) Atropine decreased the peak and TTX decreased the peak further in both the control subjects and diabetic patients. ( c ) MRS2500 increased the nadir but L-NNA abolished relaxation in the control subjects. ( d ) Also in the diabetic patients, L-NNA abolished relaxation. The Wilcoxon signed-rank test was used to evaluate the effects of each drug by compare values to the previous one.
    Figure Legend Snippet: Electrical field stimulation (EFS)-induced response of distal gastric circular muscle strips after serial administration of atropine, MRS2500, N-nitro-L-arginine (L-NNA), and tetrodotoxin (TTX). ( a and b ) Atropine decreased the peak and TTX decreased the peak further in both the control subjects and diabetic patients. ( c ) MRS2500 increased the nadir but L-NNA abolished relaxation in the control subjects. ( d ) Also in the diabetic patients, L-NNA abolished relaxation. The Wilcoxon signed-rank test was used to evaluate the effects of each drug by compare values to the previous one.

    Techniques Used:

    19) Product Images from "Subunit- and pathway-specific localization of NMDA receptors and scaffolding proteins at ganglion cell synapses in rat retina"

    Article Title: Subunit- and pathway-specific localization of NMDA receptors and scaffolding proteins at ganglion cell synapses in rat retina

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    doi: 10.1523/JNEUROSCI.5602-08.2009

    Distinct NMDAR subtype contributions to sEPSCs at ON and OFF synapses. A, Average sEPSCs recorded form a morphologically identified ON RGC (V hold = −80 mV, 1 μM TTX, 100 μM D-serine, 0 [Mg 2+ ] o , inhibition blocked). Reducing glutamate uptake with the transporter antagonist TBOA (10 μM, green, 95 events averaged) conferred a slow component onto the sEPSC waveform compared to control (black, 135 events). This TBOA-induced component was reduced by the NR2B NMDAR-specific antagonist Ro-25,6981 (Ro, 1 μM, red, 106 events) and abolished completely by the pan-NMDAR antagonist CPP (10 μM, blue, 110 events). B , Summarized effects of TBOA, Ro-25,6981 and CPP in 5 ON RGCs. C , Average sEPSCs recorded from an identified OFF RGC in the same control conditions as above (black, 425 events). In the absence of TBOA, Ro-25,6981 (red, 202 events) had no effect on the sEPSC waveform but CPP blocked a slow component (blue, 88 events), indicating the presence of synaptic NMDARs that lack NR2B subunits. D , Summarized effects of Ro-25,6981 and CPP in 5 OFF RGCs. E , Average sEPSCs recorded from an identified OFF RGC in the same control conditions as above (black, 133 events). Addition of TBOA enhanced the sEPSC waveform (green, 110 events), revealing a component that was eliminated by Ro-25,6981 (red, 120 events). CPP reduced the sEPSC waveform further (blue, 100 events). F , Summarized effects of TBOA, Ro-25,6981 and CPP in 5 OFF RGCs. G , Average sEPSCs recorded from an identified OFF RGC in the presence of TBOA (with inhibition blocked), in control (2.5 mM) [Ca 2+ ] o (black, 89 events) or low (0.5 mM) [Ca 2+ ] o (gray, 59 events). H , Effects of changing [Ca 2+ ] o on sEPSC frequency and charge transfer in 4 OFF RGCs.
    Figure Legend Snippet: Distinct NMDAR subtype contributions to sEPSCs at ON and OFF synapses. A, Average sEPSCs recorded form a morphologically identified ON RGC (V hold = −80 mV, 1 μM TTX, 100 μM D-serine, 0 [Mg 2+ ] o , inhibition blocked). Reducing glutamate uptake with the transporter antagonist TBOA (10 μM, green, 95 events averaged) conferred a slow component onto the sEPSC waveform compared to control (black, 135 events). This TBOA-induced component was reduced by the NR2B NMDAR-specific antagonist Ro-25,6981 (Ro, 1 μM, red, 106 events) and abolished completely by the pan-NMDAR antagonist CPP (10 μM, blue, 110 events). B , Summarized effects of TBOA, Ro-25,6981 and CPP in 5 ON RGCs. C , Average sEPSCs recorded from an identified OFF RGC in the same control conditions as above (black, 425 events). In the absence of TBOA, Ro-25,6981 (red, 202 events) had no effect on the sEPSC waveform but CPP blocked a slow component (blue, 88 events), indicating the presence of synaptic NMDARs that lack NR2B subunits. D , Summarized effects of Ro-25,6981 and CPP in 5 OFF RGCs. E , Average sEPSCs recorded from an identified OFF RGC in the same control conditions as above (black, 133 events). Addition of TBOA enhanced the sEPSC waveform (green, 110 events), revealing a component that was eliminated by Ro-25,6981 (red, 120 events). CPP reduced the sEPSC waveform further (blue, 100 events). F , Summarized effects of TBOA, Ro-25,6981 and CPP in 5 OFF RGCs. G , Average sEPSCs recorded from an identified OFF RGC in the presence of TBOA (with inhibition blocked), in control (2.5 mM) [Ca 2+ ] o (black, 89 events) or low (0.5 mM) [Ca 2+ ] o (gray, 59 events). H , Effects of changing [Ca 2+ ] o on sEPSC frequency and charge transfer in 4 OFF RGCs.

    Techniques Used: Inhibition

    20) Product Images from "Nitrergic Pathway Is the Major Mechanism for the Effect of DA-9701 on the Rat Gastric Fundus Relaxation"

    Article Title: Nitrergic Pathway Is the Major Mechanism for the Effect of DA-9701 on the Rat Gastric Fundus Relaxation

    Journal: Journal of Neurogastroenterology and Motility

    doi: 10.5056/jnm13098

    Electrical field stimulation-induced response of the rat gastric fundus longitudinal muscle strips after serial administration of atropine, DA-9701, N-nitro-L-arginine (L-NNA), MRS2500 and tetrodotoxin (TTX). (A, B) When atropine was added, peak and nadir did not show significant change (by Wilcoxon signed ranks test). (C, D) When DA-9701 (0.5, 5, 25 and 50 μg) was added in the presence of atropine, peak and nadir did not show significant dose-dependent change (Kruskal Wallis test for testing differences in EFS-induced contractile responses for different DA-9701 doses). (E) Subsequent addition of L-NNA, MRS2500 and TTX in the presence of atropine and DA-9701 (50 μg) did not affect peak (compared with previous value by Wilcoxon signed ranks test). (F) Subsequent addition of L-NNA in the presence of atropine and DA-9701 (50 μg) decreased nadir by inhibiting relaxation, while addition of MRS2500 and TTX in the presence of atropine, DA-9701 (50 μg) and L-NNA did not affect nadir further ( * P
    Figure Legend Snippet: Electrical field stimulation-induced response of the rat gastric fundus longitudinal muscle strips after serial administration of atropine, DA-9701, N-nitro-L-arginine (L-NNA), MRS2500 and tetrodotoxin (TTX). (A, B) When atropine was added, peak and nadir did not show significant change (by Wilcoxon signed ranks test). (C, D) When DA-9701 (0.5, 5, 25 and 50 μg) was added in the presence of atropine, peak and nadir did not show significant dose-dependent change (Kruskal Wallis test for testing differences in EFS-induced contractile responses for different DA-9701 doses). (E) Subsequent addition of L-NNA, MRS2500 and TTX in the presence of atropine and DA-9701 (50 μg) did not affect peak (compared with previous value by Wilcoxon signed ranks test). (F) Subsequent addition of L-NNA in the presence of atropine and DA-9701 (50 μg) decreased nadir by inhibiting relaxation, while addition of MRS2500 and TTX in the presence of atropine, DA-9701 (50 μg) and L-NNA did not affect nadir further ( * P

    Techniques Used:

    21) Product Images from "TRPM8 Channel Activation Reduces the Spontaneous Contractions in Human Distal Colon"

    Article Title: TRPM8 Channel Activation Reduces the Spontaneous Contractions in Human Distal Colon

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21155403

    Concentration–response curves for the inhibitory effects induced by DIPA 1–8 (1 nM–100 μM) before and after TTX (1 μM) ( A ) and TEA (10 mM) ( B ). All values are means ± S.E.M ( n = 6) and are expressed as percentage of inhibition of the spontaneous contractions. * p
    Figure Legend Snippet: Concentration–response curves for the inhibitory effects induced by DIPA 1–8 (1 nM–100 μM) before and after TTX (1 μM) ( A ) and TEA (10 mM) ( B ). All values are means ± S.E.M ( n = 6) and are expressed as percentage of inhibition of the spontaneous contractions. * p

    Techniques Used: Concentration Assay, Inhibition

    22) Product Images from "Enhanced prefrontal serotonin 5-HT1A currents in a mouse model of Williams-Beuren syndrome with low innate anxiety"

    Article Title: Enhanced prefrontal serotonin 5-HT1A currents in a mouse model of Williams-Beuren syndrome with low innate anxiety

    Journal: Journal of neurodevelopmental disorders

    doi: 10.1007/s11689-010-9044-5

    Serotonergic currents in layer V pyramidal neurons mediated by 5-HT 1A receptors in wildtype (WT) and Gtf2ird1 −/− mice. In neurons from each genotype, (1.) the 5-HT-elicited outward currents were suppressed by (2.) the selective antagonist of 5-HT 1A receptors, WAY-100635 (30 nM, 10 min). Gray bar indicates 5-HT application. All the above traces were recorded after application of TTX (2 µM, 10 min), indicating that the 5-HT outward currents were mediated directly by 5-HT 1A receptors located on the recorded cells
    Figure Legend Snippet: Serotonergic currents in layer V pyramidal neurons mediated by 5-HT 1A receptors in wildtype (WT) and Gtf2ird1 −/− mice. In neurons from each genotype, (1.) the 5-HT-elicited outward currents were suppressed by (2.) the selective antagonist of 5-HT 1A receptors, WAY-100635 (30 nM, 10 min). Gray bar indicates 5-HT application. All the above traces were recorded after application of TTX (2 µM, 10 min), indicating that the 5-HT outward currents were mediated directly by 5-HT 1A receptors located on the recorded cells

    Techniques Used: Mouse Assay

    23) Product Images from "Differential Interactions of Na+ Channel Toxins with T-type Ca2+ Channels"

    Article Title: Differential Interactions of Na+ Channel Toxins with T-type Ca2+ Channels

    Journal: The Journal of General Physiology

    doi: 10.1085/jgp.200709883

    Effects of TTX and STX on Ca V 3.3 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings obtained from HEK-293 cells transiently transfected with Ca V 3.3. This panel illustrates the effects of 750 μM Ni 2+ alone (left), those of 30 μM TTX in the absence or presence of 750 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 750 μM Ni 2+ (right). As for Figs. 6 and 7 , current traces were elicited by voltage steps to −40 mV from a holding potential of −90 mV. The three families of traces are from different cells. (B) Bar graph summarizing the effects of different blockers for experiments similar to those illustrated in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.
    Figure Legend Snippet: Effects of TTX and STX on Ca V 3.3 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings obtained from HEK-293 cells transiently transfected with Ca V 3.3. This panel illustrates the effects of 750 μM Ni 2+ alone (left), those of 30 μM TTX in the absence or presence of 750 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 750 μM Ni 2+ (right). As for Figs. 6 and 7 , current traces were elicited by voltage steps to −40 mV from a holding potential of −90 mV. The three families of traces are from different cells. (B) Bar graph summarizing the effects of different blockers for experiments similar to those illustrated in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.

    Techniques Used: Transfection, Blocking Assay

    Effects of TTX and STX on Ca V 3.2 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings showing the effects of 15 μM Ni 2+ alone (left), the effects of 30 μM TTX in the presence of 15 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 15 μM Ni 2+ (right). Please note that the leftward and middle traces were obtained from the same cell while the ones on the right hand side were from a different experiment. As for the native T-type Ca 2+ current, TTX attenuated the block of Ca V 3.1-induced current by Ni 2+ and STX produced a significant inhibition of this current. (B) Bar graph showing pooled data from similar experiments to those shown in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.
    Figure Legend Snippet: Effects of TTX and STX on Ca V 3.2 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings showing the effects of 15 μM Ni 2+ alone (left), the effects of 30 μM TTX in the presence of 15 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 15 μM Ni 2+ (right). Please note that the leftward and middle traces were obtained from the same cell while the ones on the right hand side were from a different experiment. As for the native T-type Ca 2+ current, TTX attenuated the block of Ca V 3.1-induced current by Ni 2+ and STX produced a significant inhibition of this current. (B) Bar graph showing pooled data from similar experiments to those shown in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.

    Techniques Used: Blocking Assay, Produced, Inhibition

    Effects of TTX and STX on Ca V 3.1 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings showing the effects of 550 μM Ni 2+ alone (left), the effects of 30 μM TTX in the presence of 550 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 550 μM Ni 2+ (right). Please note that the leftward and middle traces were obtained from the same cell while the ones on the righthand side were from a different experiment. As for the native T-type Ca 2+ current, TTX attenuated the block of Ca V 3.1-induced current by Ni 2+ and STX produced a significant inhibition of this current. (B) Bar graph showing pooled data from similar experiments to those shown in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.
    Figure Legend Snippet: Effects of TTX and STX on Ca V 3.1 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings showing the effects of 550 μM Ni 2+ alone (left), the effects of 30 μM TTX in the presence of 550 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 550 μM Ni 2+ (right). Please note that the leftward and middle traces were obtained from the same cell while the ones on the righthand side were from a different experiment. As for the native T-type Ca 2+ current, TTX attenuated the block of Ca V 3.1-induced current by Ni 2+ and STX produced a significant inhibition of this current. (B) Bar graph showing pooled data from similar experiments to those shown in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.

    Techniques Used: Blocking Assay, Produced, Inhibition

    24) Product Images from "Ca2+-permeable AMPA receptors in mouse olfactory bulb astrocytes"

    Article Title: Ca2+-permeable AMPA receptors in mouse olfactory bulb astrocytes

    Journal: Scientific Reports

    doi: 10.1038/srep44817

    Naspm reduces AMPA receptor-induced Ca 2+ transients and membrane currents in periglomerular astrocytes. ( a ) Effect of Naspm (50 μM) on AMPA-induced Ca 2+ transients after incubation of brain slices in bafilomycin A1 and TTX. ( b ) Naspm significantly reduced AMPA-induced Ca 2+ transients in periglomerular astrocytes. Wash out of Naspm led to a significant recovery of the Ca 2+ response. ( c ) Increasing external K + to 50 mM evoked large Ca 2+ transients in neurons, but only small Ca 2+ rises in astrocytes, indicating lack of voltage-gated Ca 2+ influx in astrocytes. ( d ) Fluo-4-loaded isolated astrocyte. Scale bar: 20 μm. ( e ) Effect of Naspm (50 μM) on AMPA-evoked Ca 2+ transients in an isolated olfactory bulb astrocyte. ( f ) Immunostaining of an eGFP-positive periglomerular astrocyte in an hGFAP-eGFP mouse (anti-GFP, green) and colabeling of GFAP as a marker for astrocytes (anti-GFAP, red). Scale bar: 10 μm. ( g ) Whole-cell current trace of a dissociated astrocyte recorded in BaCl 2 (100 μM), quinine (100 μM) and cyclothiazide (100 μM). Kainate (500 μM) evoked an inward current that was entirely blocked by GYKI 53655 (100 μM), but ( h ) was only partly reduced by Naspm (50 μM). ( i ) Normalized averaged effects of GYKI 53655and Naspm on kainate-induced currents.
    Figure Legend Snippet: Naspm reduces AMPA receptor-induced Ca 2+ transients and membrane currents in periglomerular astrocytes. ( a ) Effect of Naspm (50 μM) on AMPA-induced Ca 2+ transients after incubation of brain slices in bafilomycin A1 and TTX. ( b ) Naspm significantly reduced AMPA-induced Ca 2+ transients in periglomerular astrocytes. Wash out of Naspm led to a significant recovery of the Ca 2+ response. ( c ) Increasing external K + to 50 mM evoked large Ca 2+ transients in neurons, but only small Ca 2+ rises in astrocytes, indicating lack of voltage-gated Ca 2+ influx in astrocytes. ( d ) Fluo-4-loaded isolated astrocyte. Scale bar: 20 μm. ( e ) Effect of Naspm (50 μM) on AMPA-evoked Ca 2+ transients in an isolated olfactory bulb astrocyte. ( f ) Immunostaining of an eGFP-positive periglomerular astrocyte in an hGFAP-eGFP mouse (anti-GFP, green) and colabeling of GFAP as a marker for astrocytes (anti-GFAP, red). Scale bar: 10 μm. ( g ) Whole-cell current trace of a dissociated astrocyte recorded in BaCl 2 (100 μM), quinine (100 μM) and cyclothiazide (100 μM). Kainate (500 μM) evoked an inward current that was entirely blocked by GYKI 53655 (100 μM), but ( h ) was only partly reduced by Naspm (50 μM). ( i ) Normalized averaged effects of GYKI 53655and Naspm on kainate-induced currents.

    Techniques Used: Incubation, Isolation, Immunostaining, Marker

    25) Product Images from "Human Breast Cancer Cells Demonstrate Electrical Excitability"

    Article Title: Human Breast Cancer Cells Demonstrate Electrical Excitability

    Journal: Frontiers in Neuroscience

    doi: 10.3389/fnins.2020.00404

    VGSC activity of MDA-MB-231 cells. VGSC activity was blocked using TTX (20 μM). (A) Current trace showing electrical activity before, during and after application of TTX. (B) Quantification of the spikes recorded in (A) . Number of spikes were measured in time bins of 6 min. Bars represent average values of all 3 repeats and error bars represent SDs.
    Figure Legend Snippet: VGSC activity of MDA-MB-231 cells. VGSC activity was blocked using TTX (20 μM). (A) Current trace showing electrical activity before, during and after application of TTX. (B) Quantification of the spikes recorded in (A) . Number of spikes were measured in time bins of 6 min. Bars represent average values of all 3 repeats and error bars represent SDs.

    Techniques Used: Activity Assay, Multiple Displacement Amplification

    26) Product Images from "Divergent Roles of p75NTR and Trk Receptors in BDNF's Effects on Dendritic Spine Density and Morphology"

    Article Title: Divergent Roles of p75NTR and Trk Receptors in BDNF's Effects on Dendritic Spine Density and Morphology

    Journal: Neural Plasticity

    doi: 10.1155/2012/578057

    Role of neuronal activity in k-252a's effects on dendritic spine density and morphology. (a) Representative examples of dendritic segments of CA1 pyramidal neurons maintained in serum-containing media (SM) and treated with k-252a (200 nM), TTX (1 μ M), or both k-252a and TTX for 48 hrs (scale bar represents 2 μ m). (b) Dendritic spine density expressed per 10 μ m of apical dendrite. (c) Proportion of each morphological type of dendritic spines, expressed as a fraction of the total spine population. * P
    Figure Legend Snippet: Role of neuronal activity in k-252a's effects on dendritic spine density and morphology. (a) Representative examples of dendritic segments of CA1 pyramidal neurons maintained in serum-containing media (SM) and treated with k-252a (200 nM), TTX (1 μ M), or both k-252a and TTX for 48 hrs (scale bar represents 2 μ m). (b) Dendritic spine density expressed per 10 μ m of apical dendrite. (c) Proportion of each morphological type of dendritic spines, expressed as a fraction of the total spine population. * P

    Techniques Used: Activity Assay

    27) Product Images from "Modulation of specific intestinal epithelial progenitors by enteric neurons"

    Article Title: Modulation of specific intestinal epithelial progenitors by enteric neurons

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

    doi: 10.1073/pnas.211278098

    GLP-2 induces c-Fos-like expression in neurons and crypts. Crypt response is neuron-dependent. ( A–D ) Fluorescence micrographs of whole-mount preparations of jejunal and colonic muscle coats stained for c-Fos. Myenteric and submucosal neurons are visible in these preparations and both types respond. ( B and D ) Increased numbers of c-Fos-immunopositive nuclei in the enteric ganglia 15 min after i.v. injection of GLP-2. ( A and C ) PBS-injected controls showing background activity. In jejunum there were 38 ± 3.0 (mean ± SEM) c-Fos-positive nuclei per field in GLP-2 treated versus 12.67 ± 2.0 in controls, whereas in colon there were 242 ± 27.7 in treated versus 39.3 ± 17.9 in controls. Enteric ganglia were first identified with bright-field microscopy and then c-Fos-positive nuclei in the field were counted under fluorescence. ( E – L ) Topical TTX, a voltage-gated sodium channel blocker, inhibits the GLP-2-induced c-Fos response in crypt cells in jejunum and colon. ( E and I ) Control mice after topical PBS and an i.v. PBS injection (untreated animals are similar, not shown). In jejunum ( E ) c-Fos-positive nuclei were seen in villus epithelium but not in crypts. ( Inset ) An enlarged image of the crypt indicated by an arrow in the main figure. Colon ( I ) has positive nuclei in surface epithelium and upper crypt. ( F and J ) Control mice after topical TTX treatment and an i.v. PBS injection. In jejunum ( F ) c-Fos-positive nuclei were similarly seen in the villus but not in crypts, and surface and crypt top in colon ( J ). ( G and K ) In mice given topical PBS and an i.v. GLP-2 injection, c-Fos was expressed throughout the crypt except the base (arrowhead in Inset ) in jejunum ( G ). Colon crypts also responded ( K ). ( H and L ) Topical TTX suppresses the crypt response to an i.v. GLP-2 injection in jejunum ( H ) and colon ( L ). (Magnifications: A – D , ×30; E – H , ×70 ( Insets , ×160); I – L , ×100.)
    Figure Legend Snippet: GLP-2 induces c-Fos-like expression in neurons and crypts. Crypt response is neuron-dependent. ( A–D ) Fluorescence micrographs of whole-mount preparations of jejunal and colonic muscle coats stained for c-Fos. Myenteric and submucosal neurons are visible in these preparations and both types respond. ( B and D ) Increased numbers of c-Fos-immunopositive nuclei in the enteric ganglia 15 min after i.v. injection of GLP-2. ( A and C ) PBS-injected controls showing background activity. In jejunum there were 38 ± 3.0 (mean ± SEM) c-Fos-positive nuclei per field in GLP-2 treated versus 12.67 ± 2.0 in controls, whereas in colon there were 242 ± 27.7 in treated versus 39.3 ± 17.9 in controls. Enteric ganglia were first identified with bright-field microscopy and then c-Fos-positive nuclei in the field were counted under fluorescence. ( E – L ) Topical TTX, a voltage-gated sodium channel blocker, inhibits the GLP-2-induced c-Fos response in crypt cells in jejunum and colon. ( E and I ) Control mice after topical PBS and an i.v. PBS injection (untreated animals are similar, not shown). In jejunum ( E ) c-Fos-positive nuclei were seen in villus epithelium but not in crypts. ( Inset ) An enlarged image of the crypt indicated by an arrow in the main figure. Colon ( I ) has positive nuclei in surface epithelium and upper crypt. ( F and J ) Control mice after topical TTX treatment and an i.v. PBS injection. In jejunum ( F ) c-Fos-positive nuclei were similarly seen in the villus but not in crypts, and surface and crypt top in colon ( J ). ( G and K ) In mice given topical PBS and an i.v. GLP-2 injection, c-Fos was expressed throughout the crypt except the base (arrowhead in Inset ) in jejunum ( G ). Colon crypts also responded ( K ). ( H and L ) Topical TTX suppresses the crypt response to an i.v. GLP-2 injection in jejunum ( H ) and colon ( L ). (Magnifications: A – D , ×30; E – H , ×70 ( Insets , ×160); I – L , ×100.)

    Techniques Used: Expressing, Fluorescence, Staining, Injection, Activity Assay, Microscopy, Mouse Assay

    28) Product Images from "Regional and Developmental Differences in Na+ Currents in Vestibular Primary Afferent Neurons"

    Article Title: Regional and Developmental Differences in Na+ Currents in Vestibular Primary Afferent Neurons

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2018.00423

    A TTX-insensitive current is present in immature calyces. (A) Control I Na and currents remaining in the presence of 200 nM TTX at membrane potentials above –60 mV in a P8 calyx (note different current scales for left and right panels). Voltage protocol similar to Figure 2 : a 40 ms step to –130 mV from a holding potential of –80 mV was followed by a series of 40 ms depolarizing steps in 10 mV increments from –90 to +20 mV. (B) Control (black) and response to 200 nM TTX (red) for nine calyces (P5–11) are shown in the IV plot of peak inward currents for steps between –80 and 0 mV. (C) Left panel: the residual current following 200 nM TTX is blocked in 1 μM TTX and the block reverses with washout in a P6 calyx. Currents in response to a voltage step from –130 to –30 mV. Right panel: 1 μM JZTX-III blocks a component of I Na and the remaining current is abolished following application of 1 μM JZTX-III plus 200 nM TTX in a P7 calyx. Currents in response to a voltage step from –130 to –50 mV. (D) Summary for a group of 12 cells perfused with 200 nM TTX (six cells at P5–6, six at P8–11). A group of cells exposed to 1 μM TTX ( n = 8, one cell at P6, seven cells at P7–10) I Na was abolished in 1 μM JZTX-III and 200 nM TTX ( n = 4, two cells at P6 and two cells at P7). Peak inward current was measured at –50 mV step. I Na tended to be larger at younger ages as shown by distributions. In the presence of 200 nM TTX, I Na decreased from –3.9 (1.8) to –0.25 (0.5) nA ( P
    Figure Legend Snippet: A TTX-insensitive current is present in immature calyces. (A) Control I Na and currents remaining in the presence of 200 nM TTX at membrane potentials above –60 mV in a P8 calyx (note different current scales for left and right panels). Voltage protocol similar to Figure 2 : a 40 ms step to –130 mV from a holding potential of –80 mV was followed by a series of 40 ms depolarizing steps in 10 mV increments from –90 to +20 mV. (B) Control (black) and response to 200 nM TTX (red) for nine calyces (P5–11) are shown in the IV plot of peak inward currents for steps between –80 and 0 mV. (C) Left panel: the residual current following 200 nM TTX is blocked in 1 μM TTX and the block reverses with washout in a P6 calyx. Currents in response to a voltage step from –130 to –30 mV. Right panel: 1 μM JZTX-III blocks a component of I Na and the remaining current is abolished following application of 1 μM JZTX-III plus 200 nM TTX in a P7 calyx. Currents in response to a voltage step from –130 to –50 mV. (D) Summary for a group of 12 cells perfused with 200 nM TTX (six cells at P5–6, six at P8–11). A group of cells exposed to 1 μM TTX ( n = 8, one cell at P6, seven cells at P7–10) I Na was abolished in 1 μM JZTX-III and 200 nM TTX ( n = 4, two cells at P6 and two cells at P7). Peak inward current was measured at –50 mV step. I Na tended to be larger at younger ages as shown by distributions. In the presence of 200 nM TTX, I Na decreased from –3.9 (1.8) to –0.25 (0.5) nA ( P

    Techniques Used: Mass Spectrometry, Blocking Assay

    29) Product Images from "Identification of Persistent and Resurgent Sodium Currents in Spiral Ganglion Neurons Cultured from the Mouse Cochlea"

    Article Title: Identification of Persistent and Resurgent Sodium Currents in Spiral Ganglion Neurons Cultured from the Mouse Cochlea

    Journal: eNeuro

    doi: 10.1523/ENEURO.0303-17.2017

    TTX-sensitive transient and persistent Na + currents identified in SGNs cultured from hearing mice. A , Current responses recorded using K + -filled patch electrodes, from a representative rapidly-adapting P14 SGN during 200-ms voltage steps. A transient inward Na + current (I NaT ) was followed by LVA and HVA outward K + currents. B , Current response from the same cell during a voltage ramp (between −143 and +47 mV, duration 400 ms) before (control) and after bath application of 100 nM DTX-K. DTX-K blocked the LVA K + current, revealing a small inward current activated at potentials positive to −65 mV (detailed in the inset). C , Current responses recorded using K + -filled patch electrodes, from a representative nonadapting P14 SGN during 200-ms voltage steps (protocol described in A ). Activation of I NaT was followed by activation of HVA K + currents. LVA K + currents were absent from this cell. D , Current responses from the same SGN, during a voltage ramp before (control) and after bath application of 100 nM TTX. A TTX-sensitive inward current activated at potentials positive to −65 mV (difference current, detailed in the inset). E , TTX-subtracted currents recorded from the SGN in D , during 200-ms depolarizing voltage steps. A voltage step to −53 mV activated a small persistent inward current (I NaP ), whereas the voltage step to −33 mV activated a large I NaT ( * , peak current cropped for clarity) and a small I NaP . F , Comparison of the voltage dependence of TTX-sensitive I NaT and I NaP (measured at the arrow in E ) in this SGN.
    Figure Legend Snippet: TTX-sensitive transient and persistent Na + currents identified in SGNs cultured from hearing mice. A , Current responses recorded using K + -filled patch electrodes, from a representative rapidly-adapting P14 SGN during 200-ms voltage steps. A transient inward Na + current (I NaT ) was followed by LVA and HVA outward K + currents. B , Current response from the same cell during a voltage ramp (between −143 and +47 mV, duration 400 ms) before (control) and after bath application of 100 nM DTX-K. DTX-K blocked the LVA K + current, revealing a small inward current activated at potentials positive to −65 mV (detailed in the inset). C , Current responses recorded using K + -filled patch electrodes, from a representative nonadapting P14 SGN during 200-ms voltage steps (protocol described in A ). Activation of I NaT was followed by activation of HVA K + currents. LVA K + currents were absent from this cell. D , Current responses from the same SGN, during a voltage ramp before (control) and after bath application of 100 nM TTX. A TTX-sensitive inward current activated at potentials positive to −65 mV (difference current, detailed in the inset). E , TTX-subtracted currents recorded from the SGN in D , during 200-ms depolarizing voltage steps. A voltage step to −53 mV activated a small persistent inward current (I NaP ), whereas the voltage step to −33 mV activated a large I NaT ( * , peak current cropped for clarity) and a small I NaP . F , Comparison of the voltage dependence of TTX-sensitive I NaT and I NaP (measured at the arrow in E ) in this SGN.

    Techniques Used: Cell Culture, Mouse Assay, Mass Spectrometry, Activation Assay

    30) Product Images from "Expression of TRPV1 channels by Cajal‐Retzius cells and layer‐specific modulation of synaptic transmission by capsaicin in the mouse hippocampus"

    Article Title: Expression of TRPV1 channels by Cajal‐Retzius cells and layer‐specific modulation of synaptic transmission by capsaicin in the mouse hippocampus

    Journal: The Journal of Physiology

    doi: 10.1113/JP275685

    Responses to capsaicin are not dependent on the integrity of synaptic transmission and/or action potential generation A , left panel, control image in the presence of antagonists of synaptic ionotropic receptors (syn blockers; NBQX, 20 μM; D‐AP5, 50 μM; picrotoxinin, 50 μM) and after the addition of capsaicin (1 μM). Middle panel, similar to left panel with the additional constant presence of TTX. Right panel, control image in the presence of TTX and cadmium (150 μM) and after the introduction of capsaicin (1 μM). Notice that none of the experimental conditions was successful in preventing responses to capsaicin. Colour scale bar in arbitrary units. B , F / F 0 traces obtained from several CRs shown in different colours and superimposed in the same experimental conditions of the panels in A . C , summary plots from several experiments quantifying the peak of the F / F 0 response (left), its half‐width (middle) and latency (right).
    Figure Legend Snippet: Responses to capsaicin are not dependent on the integrity of synaptic transmission and/or action potential generation A , left panel, control image in the presence of antagonists of synaptic ionotropic receptors (syn blockers; NBQX, 20 μM; D‐AP5, 50 μM; picrotoxinin, 50 μM) and after the addition of capsaicin (1 μM). Middle panel, similar to left panel with the additional constant presence of TTX. Right panel, control image in the presence of TTX and cadmium (150 μM) and after the introduction of capsaicin (1 μM). Notice that none of the experimental conditions was successful in preventing responses to capsaicin. Colour scale bar in arbitrary units. B , F / F 0 traces obtained from several CRs shown in different colours and superimposed in the same experimental conditions of the panels in A . C , summary plots from several experiments quantifying the peak of the F / F 0 response (left), its half‐width (middle) and latency (right).

    Techniques Used: Transmission Assay

    31) Product Images from "Presynaptic NMDARs cooperate with local spikes toward GABA release from the reciprocal olfactory bulb granule cell spine"

    Article Title: Presynaptic NMDARs cooperate with local spikes toward GABA release from the reciprocal olfactory bulb granule cell spine

    Journal: eLife

    doi: 10.7554/eLife.63737

    Effects of Na v blockade (TTX, 500 nM) on spontaneous activity. ( A ) Representative experiment showing voltage-clamp recordings (+10 mV) from a mitral cell in the presence of DNI (control, top trace), upon addition of TTX (middle trace) and further wash-in of BCC (50 µM; bottom trace). ( B ) sIPSCs in control and in presence of TTX (n = 10 MCs). Left: Frequency. Right: Amplitude. sIPSCs were abolished in BCC (analysis not shown) ( C ) Linear regression between the TTX effect on sIPSC frequency vs urIPSC amplitude (n = 10 MCs). No significant correlation.
    Figure Legend Snippet: Effects of Na v blockade (TTX, 500 nM) on spontaneous activity. ( A ) Representative experiment showing voltage-clamp recordings (+10 mV) from a mitral cell in the presence of DNI (control, top trace), upon addition of TTX (middle trace) and further wash-in of BCC (50 µM; bottom trace). ( B ) sIPSCs in control and in presence of TTX (n = 10 MCs). Left: Frequency. Right: Amplitude. sIPSCs were abolished in BCC (analysis not shown) ( C ) Linear regression between the TTX effect on sIPSC frequency vs urIPSC amplitude (n = 10 MCs). No significant correlation.

    Techniques Used: Activity Assay

    32) Product Images from "Corticotropin-releasing factor increases Purkinje neuron excitability by modulating sodium, potassium, and Ih currents"

    Article Title: Corticotropin-releasing factor increases Purkinje neuron excitability by modulating sodium, potassium, and Ih currents

    Journal: Journal of Neurophysiology

    doi: 10.1152/jn.00745.2015

    CRF elicits voltage changes in PN. A : the average of 4 responses of PN to local application of CRF (8 μM, red bar) at four different holding potentials in the presence of tetrodotoxin (TTX). Note the reversal of the response between −40 mV and −50 mV. B : ZD-7288 blocks the TTX-insensitive depolarizing response. The integral of the depolarizing response measured from CRF onset over a duration of 5.5 s under control conditions ( n = 21), in the presence of TTX ( n = 13) and TTX + ZD-7288 ( n = 15) is shown. C , first trace: whole cell recording of the average response to CRF application in the presence of TTX. Second trace: 8 pulses of −40 pA for 0.5 s, delivered at 1 Hz during CRF response. The third trace is the subtraction of the first trace from the second trace. Red bar denote the CRF application. Note the reduction in the response to current injections during CRF application. Fourth trace is the current injection protocol. D : the average reduction in input resistance during CRF application in 5 PN. E : a representative example of seven superimposed traces of the hyperpolarizing response of PN to CRF applications obtained from different holding potentials (−75 to −35 mV) and aligned by the membrane voltage before the application. F : the result shown in E plotted as a function of the membrane potential (red curve) and similar curves measured from another 5 cells (black curves). 2 μM CRF was used in C – F .
    Figure Legend Snippet: CRF elicits voltage changes in PN. A : the average of 4 responses of PN to local application of CRF (8 μM, red bar) at four different holding potentials in the presence of tetrodotoxin (TTX). Note the reversal of the response between −40 mV and −50 mV. B : ZD-7288 blocks the TTX-insensitive depolarizing response. The integral of the depolarizing response measured from CRF onset over a duration of 5.5 s under control conditions ( n = 21), in the presence of TTX ( n = 13) and TTX + ZD-7288 ( n = 15) is shown. C , first trace: whole cell recording of the average response to CRF application in the presence of TTX. Second trace: 8 pulses of −40 pA for 0.5 s, delivered at 1 Hz during CRF response. The third trace is the subtraction of the first trace from the second trace. Red bar denote the CRF application. Note the reduction in the response to current injections during CRF application. Fourth trace is the current injection protocol. D : the average reduction in input resistance during CRF application in 5 PN. E : a representative example of seven superimposed traces of the hyperpolarizing response of PN to CRF applications obtained from different holding potentials (−75 to −35 mV) and aligned by the membrane voltage before the application. F : the result shown in E plotted as a function of the membrane potential (red curve) and similar curves measured from another 5 cells (black curves). 2 μM CRF was used in C – F .

    Techniques Used: Injection

    33) Product Images from "Functional segregation of voltage-activated calcium channels in motoneurons of the dorsal motor nucleus of the vagus"

    Article Title: Functional segregation of voltage-activated calcium channels in motoneurons of the dorsal motor nucleus of the vagus

    Journal: Journal of Neurophysiology

    doi: 10.1152/jn.00432.2014

    A-type Kv4 K + currents slow pacemaking of DMV neurons. A , left : example of measurements of the activation (black) and inactivation (gray) of the transient K + current in the presence of TTX. A , right : normalized Boltzman sigmoidal functions describing the voltage dependence of activation and inactivation of the Kv4 current, and the distribution of the voltages of half activation ( n = 13) and inactivation ( n = 15), reveal a window Kv4 current that spans the typical subthreshold range of the voltage trajectory of DMV neurons during pacemaking. B , left : Time course of the reduction in the peak A current as a function of time after membrane rupture in whole cell configuration. B , right : comparison of the peak A current at membrane rupture vs. past 4 min from rupture. C , left : time course of the instantaneous firing rate of a DMV neuron in cell-attached mode before membrane rupture and after whole cell configuration was achieved. B , right : comparison of autonomous discharge rate before and after rupturing the membrane. D : brief application of nominally 1–2 μM phrixotoxin-2 (PaTX-2), a selective Kv4 antagonist, transiently increases the firing rate of DMV neurons recorded in the loose-patch configuration (shown on a logarithmic scale). * P
    Figure Legend Snippet: A-type Kv4 K + currents slow pacemaking of DMV neurons. A , left : example of measurements of the activation (black) and inactivation (gray) of the transient K + current in the presence of TTX. A , right : normalized Boltzman sigmoidal functions describing the voltage dependence of activation and inactivation of the Kv4 current, and the distribution of the voltages of half activation ( n = 13) and inactivation ( n = 15), reveal a window Kv4 current that spans the typical subthreshold range of the voltage trajectory of DMV neurons during pacemaking. B , left : Time course of the reduction in the peak A current as a function of time after membrane rupture in whole cell configuration. B , right : comparison of the peak A current at membrane rupture vs. past 4 min from rupture. C , left : time course of the instantaneous firing rate of a DMV neuron in cell-attached mode before membrane rupture and after whole cell configuration was achieved. B , right : comparison of autonomous discharge rate before and after rupturing the membrane. D : brief application of nominally 1–2 μM phrixotoxin-2 (PaTX-2), a selective Kv4 antagonist, transiently increases the firing rate of DMV neurons recorded in the loose-patch configuration (shown on a logarithmic scale). * P

    Techniques Used: Activation Assay

    34) Product Images from "Differential Interactions of Na+ Channel Toxins with T-type Ca2+ Channels"

    Article Title: Differential Interactions of Na+ Channel Toxins with T-type Ca2+ Channels

    Journal: The Journal of General Physiology

    doi: 10.1085/jgp.200709883

    Effects of TTX and STX on Ca V 3.3 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings obtained from HEK-293 cells transiently transfected with Ca V 3.3. This panel illustrates the effects of 750 μM Ni 2+ alone (left), those of 30 μM TTX in the absence or presence of 750 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 750 μM Ni 2+ (right). As for Figs. 6 and 7 , current traces were elicited by voltage steps to −40 mV from a holding potential of −90 mV. The three families of traces are from different cells. (B) Bar graph summarizing the effects of different blockers for experiments similar to those illustrated in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.
    Figure Legend Snippet: Effects of TTX and STX on Ca V 3.3 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings obtained from HEK-293 cells transiently transfected with Ca V 3.3. This panel illustrates the effects of 750 μM Ni 2+ alone (left), those of 30 μM TTX in the absence or presence of 750 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 750 μM Ni 2+ (right). As for Figs. 6 and 7 , current traces were elicited by voltage steps to −40 mV from a holding potential of −90 mV. The three families of traces are from different cells. (B) Bar graph summarizing the effects of different blockers for experiments similar to those illustrated in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.

    Techniques Used: Transfection, Blocking Assay

    Effects of TTX and STX on Ca V 3.2 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings showing the effects of 15 μM Ni 2+ alone (left), the effects of 30 μM TTX in the presence of 15 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 15 μM Ni 2+ (right). Please note that the leftward and middle traces were obtained from the same cell while the ones on the right hand side were from a different experiment. As for the native T-type Ca 2+ current, TTX attenuated the block of Ca V 3.1-induced current by Ni 2+ and STX produced a significant inhibition of this current. (B) Bar graph showing pooled data from similar experiments to those shown in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.
    Figure Legend Snippet: Effects of TTX and STX on Ca V 3.2 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings showing the effects of 15 μM Ni 2+ alone (left), the effects of 30 μM TTX in the presence of 15 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 15 μM Ni 2+ (right). Please note that the leftward and middle traces were obtained from the same cell while the ones on the right hand side were from a different experiment. As for the native T-type Ca 2+ current, TTX attenuated the block of Ca V 3.1-induced current by Ni 2+ and STX produced a significant inhibition of this current. (B) Bar graph showing pooled data from similar experiments to those shown in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.

    Techniques Used: Blocking Assay, Produced, Inhibition

    Effects of TTX and STX on Ca V 3.1 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings showing the effects of 550 μM Ni 2+ alone (left), the effects of 30 μM TTX in the presence of 550 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 550 μM Ni 2+ (right). Please note that the leftward and middle traces were obtained from the same cell while the ones on the righthand side were from a different experiment. As for the native T-type Ca 2+ current, TTX attenuated the block of Ca V 3.1-induced current by Ni 2+ and STX produced a significant inhibition of this current. (B) Bar graph showing pooled data from similar experiments to those shown in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.
    Figure Legend Snippet: Effects of TTX and STX on Ca V 3.1 expressed in HEK-293 cells. (A) Typical T-type Ca 2+ current recordings showing the effects of 550 μM Ni 2+ alone (left), the effects of 30 μM TTX in the presence of 550 μM Ni 2+ (middle), and the effects of 1 μM STX in the absence or presence of 550 μM Ni 2+ (right). Please note that the leftward and middle traces were obtained from the same cell while the ones on the righthand side were from a different experiment. As for the native T-type Ca 2+ current, TTX attenuated the block of Ca V 3.1-induced current by Ni 2+ and STX produced a significant inhibition of this current. (B) Bar graph showing pooled data from similar experiments to those shown in A. The data were expressed as mean ± SEM % block of peak inward current. The numbers in parentheses reflect the number of experiments. TTX and STX for all these experiments were respectively purchased from Alomone Laboratories and the Institute for Marine Biosciences, NRC-IMB.

    Techniques Used: Blocking Assay, Produced, Inhibition

    35) Product Images from "Hair cell-type dependent expression of basolateral ion channels shapes response dynamics in the frog utricle"

    Article Title: Hair cell-type dependent expression of basolateral ion channels shapes response dynamics in the frog utricle

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2015.00338

    Voltage dependent Ca currents . (A) Currents evoked in intra-Cs, extra-Ba solutions with 100 nM TTX. Note the presence of voltage-dependent inactivation. (B) Effects of Cd 200 μM (left) and nimodipine 10 μM (right). Same solutions as (A) . (C) Ca-dependent inactivation. 500 ms conditioning steps at variable potentials were followed by a test step. In the presence of 5 mM Ca (red traces), inactivation of the test step was maximal for potentials that evoked the largest current in the conditioning step. In the presence of Ba 5 mM (black traces) Ca-dependent inactivation was abolished. (D) Voltage-dependence of Ca-dependent inactivation fraction ( n = 5, upper panel) and timecourse (lower panel) (E) I-V plot for normalized inward currents in eB type hair cells ( n = 10). Filled circles, Na currents; Open circles, steady-state Ca currents from the same cells. Ca currents are obtained with protocols as in (A) , from a holding potential of –60 mV; Na currents are obtained in the presence of Cd or as subtraction between currents from holding potentials of −100 and −60 mV.
    Figure Legend Snippet: Voltage dependent Ca currents . (A) Currents evoked in intra-Cs, extra-Ba solutions with 100 nM TTX. Note the presence of voltage-dependent inactivation. (B) Effects of Cd 200 μM (left) and nimodipine 10 μM (right). Same solutions as (A) . (C) Ca-dependent inactivation. 500 ms conditioning steps at variable potentials were followed by a test step. In the presence of 5 mM Ca (red traces), inactivation of the test step was maximal for potentials that evoked the largest current in the conditioning step. In the presence of Ba 5 mM (black traces) Ca-dependent inactivation was abolished. (D) Voltage-dependence of Ca-dependent inactivation fraction ( n = 5, upper panel) and timecourse (lower panel) (E) I-V plot for normalized inward currents in eB type hair cells ( n = 10). Filled circles, Na currents; Open circles, steady-state Ca currents from the same cells. Ca currents are obtained with protocols as in (A) , from a holding potential of –60 mV; Na currents are obtained in the presence of Cd or as subtraction between currents from holding potentials of −100 and −60 mV.

    Techniques Used:

    36) Product Images from "Essential Role of Somatic Kv2 Channels in High-Frequency Firing in Cartwheel Cells of the Dorsal Cochlear Nucleus"

    Article Title: Essential Role of Somatic Kv2 Channels in High-Frequency Firing in Cartwheel Cells of the Dorsal Cochlear Nucleus

    Journal: eNeuro

    doi: 10.1523/ENEURO.0515-20.2021

    Biophysical properties of GxTX-sensitive Kv2 current in cartwheel cells. A , Outward current evoked by voltage steps. Pulse protocol is indicated at the bottom of A . The recordings were made at room temperature (23–24°C) using ACSF supplemented with NBQX, MK-801, strychnine, picrotoxin, TTX, apamin (a SK channel blocker), and penitrem A (a BK channel blocker). To remove inward current by voltage-gated calcium channels, CaCl 2 in the ACSF was excluded and replaced with equimolar MgCl 2 , and 0.25 m m EGTA-Na was added. GxTX-sensitive current was obtained by subtraction. B , The current–voltage relationship of the outward current in the absence (control) or presence of 100 n m GxTX (GxTX). The amplitude of steady-state was used for the plotting. Here and the following figures, error bars indicate SEM, numbers in parentheses indicate the number of replications (cells). Statistical significance was tested using two-way repeated measure ANOVA and Bonferroni post hoc tests (significance at p
    Figure Legend Snippet: Biophysical properties of GxTX-sensitive Kv2 current in cartwheel cells. A , Outward current evoked by voltage steps. Pulse protocol is indicated at the bottom of A . The recordings were made at room temperature (23–24°C) using ACSF supplemented with NBQX, MK-801, strychnine, picrotoxin, TTX, apamin (a SK channel blocker), and penitrem A (a BK channel blocker). To remove inward current by voltage-gated calcium channels, CaCl 2 in the ACSF was excluded and replaced with equimolar MgCl 2 , and 0.25 m m EGTA-Na was added. GxTX-sensitive current was obtained by subtraction. B , The current–voltage relationship of the outward current in the absence (control) or presence of 100 n m GxTX (GxTX). The amplitude of steady-state was used for the plotting. Here and the following figures, error bars indicate SEM, numbers in parentheses indicate the number of replications (cells). Statistical significance was tested using two-way repeated measure ANOVA and Bonferroni post hoc tests (significance at p

    Techniques Used:

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    Alomone Labs ttx tetrodotoxin
    Activity dependence of AMPAR redistribution dynamics. A ) Distribution of range over mean values for SEpH:GluA2 puncta in the presence of <t>CNQX</t> (10 µM), AP-5 (50 µM) and <t>TTX</t> (1 µM) (Nlgn-1 KO: 10 neurons, 481 synapses; WT: 10 neurons, 514 synapses). B ) SI decay rates in the presence of the aforementioned pharmacological agents.
    Ttx Tetrodotoxin, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effects of barbaloin on <t>ATX</t> II-induced late sodium current ( I Na.L ) enhancements and under normal conditions in rabbit ventricular myocytes. (A) Representative single traces of the effects of 4 μmol/L <t>TTX</t> on ATX II-induced I Na.L enhancement. (B) Representative single traces showing that barbaloin exerted reversible inhibitory effects on ATX II-induced I Na.L enhancement. (C) Representative single traces of the effects of 200 μmol/L barbaloin on I Na.L under normal conditions. (D) Single traces of the effects of barbaloin on ATX II-induced I Na.L enhancements at a membrane potential of −20 mV. (E) Representative traces of the effects of barbaloin on ATX II-induced I Na.L enhancements at membrane potentials of −80, −60, −50, −40 and −20 mV. (F) Bar graphs showing the mean ATX II-increased I Na.L percentage values for the control, 100 μmol/L barbaloin-treated and wash-out groups ( n =10. ** P
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    Activity dependence of AMPAR redistribution dynamics. A ) Distribution of range over mean values for SEpH:GluA2 puncta in the presence of CNQX (10 µM), AP-5 (50 µM) and TTX (1 µM) (Nlgn-1 KO: 10 neurons, 481 synapses; WT: 10 neurons, 514 synapses). B ) SI decay rates in the presence of the aforementioned pharmacological agents.

    Journal: PLoS ONE

    Article Title: Neuroligin-1 Loss Is Associated with Reduced Tenacity of Excitatory Synapses

    doi: 10.1371/journal.pone.0042314

    Figure Lengend Snippet: Activity dependence of AMPAR redistribution dynamics. A ) Distribution of range over mean values for SEpH:GluA2 puncta in the presence of CNQX (10 µM), AP-5 (50 µM) and TTX (1 µM) (Nlgn-1 KO: 10 neurons, 481 synapses; WT: 10 neurons, 514 synapses). B ) SI decay rates in the presence of the aforementioned pharmacological agents.

    Article Snippet: Pharmacological manipulations Reagents were procured from the following sources: CNQX (6-cyano-7 nitroquinoxaline-2,3-dione) from Tocris Bioscience; AP-5 (2-amino-5-phosphonopentanoic acid) from Sigma-Aldrich; TTX (tetrodotoxin) from Alomone Labs.

    Techniques: Activity Assay

    Effects of veratridine on human sperm motility in the presence of tetrodotoxin, A-803467 or ab-66743. The effects of veratridine (10 μM) after different incubation times were analyzed in the presence of (A) the VGSC inhibitor tetrodotoxin (TTX) (10 nM) (B) TTX (10 μM), (C) the selective Na v 1.8 antagonist A-803467 (10 μM), (D) the Na v 1.8 antibody ab-66743 (dilution 1:50) or the corresponding solvent. Bars are means with SEM of 6-8 different experiments and represent percentage changes in progressive motility (grade A+B sperm) relative to the value observed at the same time in the respective solvent-treated paired controls. * P

    Journal: PLoS ONE

    Article Title: The Voltage-Gated Sodium Channel Nav1.8 Is Expressed in Human Sperm

    doi: 10.1371/journal.pone.0076084

    Figure Lengend Snippet: Effects of veratridine on human sperm motility in the presence of tetrodotoxin, A-803467 or ab-66743. The effects of veratridine (10 μM) after different incubation times were analyzed in the presence of (A) the VGSC inhibitor tetrodotoxin (TTX) (10 nM) (B) TTX (10 μM), (C) the selective Na v 1.8 antagonist A-803467 (10 μM), (D) the Na v 1.8 antibody ab-66743 (dilution 1:50) or the corresponding solvent. Bars are means with SEM of 6-8 different experiments and represent percentage changes in progressive motility (grade A+B sperm) relative to the value observed at the same time in the respective solvent-treated paired controls. * P

    Article Snippet: In parallel experiments, the effect of veratridine or its solvent was investigated in aliquots pretreated during the last 15 min of capacitation with the specific Na v 1.8 antagonist A-803467 (Sigma) (1 μM) [ ] or during capacitation (2 h) with TTX (Alomone Labs, Jerusalem, Israel)) (0.01 or 10 μM), the Na v 1.8 antibody ab-66743 (dilution 1:50) or the corresponding solvent.

    Techniques: Incubation

    A18a/CLI2 and aCC are monosynaptically connected. (A-B) Excitatory synaptic inputs to aCC recorded in absence (A) and presence of TTX (B and inset). Four different Gal4 lines were tested to drive ChR and NaChBac expression in the cholinergic lateral interneurons 1 and 2 (CLI 1 and 2). (C) Quantification of aCC synaptic drive revealed that CLI2, but not CLI 1, is monosynaptically connected to aCC. The TTX-induced reduction of synaptic current amplitudes (e.g. CLIs-Gal4 ) suggests the existence of additional unknown intermediary neurons located upstream of aCC and activated by neurons expressing ChR; NaChBac.

    Journal: bioRxiv

    Article Title: Electrophysiological validation of premotor interneurons monosynaptically connected to the aCC motoneuron in the Drosophila larval CNS

    doi: 10.1101/2020.06.17.156430

    Figure Lengend Snippet: A18a/CLI2 and aCC are monosynaptically connected. (A-B) Excitatory synaptic inputs to aCC recorded in absence (A) and presence of TTX (B and inset). Four different Gal4 lines were tested to drive ChR and NaChBac expression in the cholinergic lateral interneurons 1 and 2 (CLI 1 and 2). (C) Quantification of aCC synaptic drive revealed that CLI2, but not CLI 1, is monosynaptically connected to aCC. The TTX-induced reduction of synaptic current amplitudes (e.g. CLIs-Gal4 ) suggests the existence of additional unknown intermediary neurons located upstream of aCC and activated by neurons expressing ChR; NaChBac.

    Article Snippet: The CLI-Gal4 line, less specific compared to the other ones, exhibited the largest synaptic drive to aCC, which is notably reduced, but not fully blocked, by the presence of TTX (−9.55 ± 3.19 pA/pF, n = 4, to -3.57 ± 0.92 pA/pF, n = 6, ).

    Techniques: Expressing

    A31k and aCC are monosynaptically connected. (A-B) Optogenetic activation of R20A03-AD; R87H09-DBD split Gal4, did not produce detectable changes in aCC firing (evoked by current injection) recorded at both L3 (A: F (2, 6) = 1.733, P = 0.3134, repeated measures one-way ANOVA, n = 3, black lines), and L1 (B: F (2, 12) = 0.5404, P = 0.5928, repeated measures one-way ANOVA, n = 5, black lines). Average values are shown in red. (C) Optogenetic activation of R20A03-AD; R93B07-DBD split Gal4 significantly reduced aCC firing (F (2, 12) = 20.22, P = 0.0011, repeated measures one-way ANOVA, n = 5, black lines). Average values are shown in red. (D) Quantification of the synaptic drive to aCC (held at -40 mV) following optogenetic activation of R20A03-AD; R93B07-DBD split Gal4, in absence or presence of 2 µM TTX. (E) Sample traces showing the optogenetic activation of R20A03-AD; R93B07-DBD split Gal4. aCC neurons were recorded both in voltage (both at -60 and -40 mV) and in current clamp (at -40 mV) in presence of TTX. (F-G) Sample traces confirming the GABAergic connection between A31k and aCC. Cells were recorded, as previously described, before (black trace) and after (gray trace) the bath application of 10 µM Picrotoxin (F), or 1 mM Gabazine (G). In both cases, aCC inputs were abolished.

    Journal: bioRxiv

    Article Title: Electrophysiological validation of premotor interneurons monosynaptically connected to the aCC motoneuron in the Drosophila larval CNS

    doi: 10.1101/2020.06.17.156430

    Figure Lengend Snippet: A31k and aCC are monosynaptically connected. (A-B) Optogenetic activation of R20A03-AD; R87H09-DBD split Gal4, did not produce detectable changes in aCC firing (evoked by current injection) recorded at both L3 (A: F (2, 6) = 1.733, P = 0.3134, repeated measures one-way ANOVA, n = 3, black lines), and L1 (B: F (2, 12) = 0.5404, P = 0.5928, repeated measures one-way ANOVA, n = 5, black lines). Average values are shown in red. (C) Optogenetic activation of R20A03-AD; R93B07-DBD split Gal4 significantly reduced aCC firing (F (2, 12) = 20.22, P = 0.0011, repeated measures one-way ANOVA, n = 5, black lines). Average values are shown in red. (D) Quantification of the synaptic drive to aCC (held at -40 mV) following optogenetic activation of R20A03-AD; R93B07-DBD split Gal4, in absence or presence of 2 µM TTX. (E) Sample traces showing the optogenetic activation of R20A03-AD; R93B07-DBD split Gal4. aCC neurons were recorded both in voltage (both at -60 and -40 mV) and in current clamp (at -40 mV) in presence of TTX. (F-G) Sample traces confirming the GABAergic connection between A31k and aCC. Cells were recorded, as previously described, before (black trace) and after (gray trace) the bath application of 10 µM Picrotoxin (F), or 1 mM Gabazine (G). In both cases, aCC inputs were abolished.

    Article Snippet: The CLI-Gal4 line, less specific compared to the other ones, exhibited the largest synaptic drive to aCC, which is notably reduced, but not fully blocked, by the presence of TTX (−9.55 ± 3.19 pA/pF, n = 4, to -3.57 ± 0.92 pA/pF, n = 6, ).

    Techniques: Activation Assay, Injection

    A23a and aCC are monosynaptically connected. (A) Optogenetic activation of R78F07-Gal4 driving ChR; NaChBac reduces action potential firing in aCC (elicited by injection of constant current). On average, we observed an inhibitory effect (F (2, 15) = 5.005, P = 0.0216, repeated measures one-way ANOVA, n = 6, black lines). Average values are shown in red. (B) Quantification of the synaptic inputs recorded from aCC (held at -40 mV) following optogenetic activation of R78F07-Gal4 driving ChR; NaChBac, before and after 2 µM TTX application. In the presence of TTX, we observed a heterogeneous range of inputs with excitation prevailing over inhibition, thus suggesting a poor specificity for this line to target the GABAergic A23a interneuron. Some recordings (2 out of 8 cells) showed a biphasic connection where both the excitatory and inhibitory components were observed in the same cell (values highlighted with a different colour, +TTX group). (C) Raw electrophysiological sweeps from an example of a biphasic connection obtained with optogenetic activation of R78F07-Gal4 . The same cell was recorded 5 times during optogenetic stimulation before (black traces) and after (red traces) TTX exposure. Whilst the inhibitory component seems to prevail before applying TTX, isolation of NaChBac -overexpressing neurons resulted in a reliable excitatory component (arrowhead) followed by a delayed erratic inhibitory component (arrow). (D) Optogenetic activation of R78F07-AD; R49C08-DBD split Gal4 did not affect aCC firing (F (2, 21) = 0.8322, P = 0.4005, repeated measures one-way ANOVA, n = 8, black lines). Average values are shown in red. (E) Optogenetic activation of R41G07-AD; R78F07-DBD split Gal4 significantly reduced aCC firing (F (2, 21) = 9.662, P = 0.0141, repeated measures one-way ANOVA, n = 8, black lines). Average values are shown in red. (F) Quantification of the synaptic drive to aCC (held at -40 mV) following optogenetic activation of R41G07-AD; R78F07-DBD split Gal4 in the absence, or presence, of 2 µM TTX. The prevalence of inhibitory inputs suggests a better specificity for this line in targeting A23a compared to the previous tested lines. (G) Sample traces showing the optogenetic activation of R41G07-AD; R78F07-DBD split Gal4, driving ChR; NaChBac. aCC were recorded both in voltage-(both at -60 and -40 mV) and in current clamp (at -40 mV) in presence of TTX. (H-I) Sample traces confirming that the A23a→ aCC synapse is GABAergic. Cells were recorded, as previously described, before (black trace) and after (gray trace) bath application of 10 µM Picrotoxin (H) or 1 mM Gabazine (I), two blockers of the Drosophila GABA A receptor. In both cases, aCC inputs were abolished.

    Journal: bioRxiv

    Article Title: Electrophysiological validation of premotor interneurons monosynaptically connected to the aCC motoneuron in the Drosophila larval CNS

    doi: 10.1101/2020.06.17.156430

    Figure Lengend Snippet: A23a and aCC are monosynaptically connected. (A) Optogenetic activation of R78F07-Gal4 driving ChR; NaChBac reduces action potential firing in aCC (elicited by injection of constant current). On average, we observed an inhibitory effect (F (2, 15) = 5.005, P = 0.0216, repeated measures one-way ANOVA, n = 6, black lines). Average values are shown in red. (B) Quantification of the synaptic inputs recorded from aCC (held at -40 mV) following optogenetic activation of R78F07-Gal4 driving ChR; NaChBac, before and after 2 µM TTX application. In the presence of TTX, we observed a heterogeneous range of inputs with excitation prevailing over inhibition, thus suggesting a poor specificity for this line to target the GABAergic A23a interneuron. Some recordings (2 out of 8 cells) showed a biphasic connection where both the excitatory and inhibitory components were observed in the same cell (values highlighted with a different colour, +TTX group). (C) Raw electrophysiological sweeps from an example of a biphasic connection obtained with optogenetic activation of R78F07-Gal4 . The same cell was recorded 5 times during optogenetic stimulation before (black traces) and after (red traces) TTX exposure. Whilst the inhibitory component seems to prevail before applying TTX, isolation of NaChBac -overexpressing neurons resulted in a reliable excitatory component (arrowhead) followed by a delayed erratic inhibitory component (arrow). (D) Optogenetic activation of R78F07-AD; R49C08-DBD split Gal4 did not affect aCC firing (F (2, 21) = 0.8322, P = 0.4005, repeated measures one-way ANOVA, n = 8, black lines). Average values are shown in red. (E) Optogenetic activation of R41G07-AD; R78F07-DBD split Gal4 significantly reduced aCC firing (F (2, 21) = 9.662, P = 0.0141, repeated measures one-way ANOVA, n = 8, black lines). Average values are shown in red. (F) Quantification of the synaptic drive to aCC (held at -40 mV) following optogenetic activation of R41G07-AD; R78F07-DBD split Gal4 in the absence, or presence, of 2 µM TTX. The prevalence of inhibitory inputs suggests a better specificity for this line in targeting A23a compared to the previous tested lines. (G) Sample traces showing the optogenetic activation of R41G07-AD; R78F07-DBD split Gal4, driving ChR; NaChBac. aCC were recorded both in voltage-(both at -60 and -40 mV) and in current clamp (at -40 mV) in presence of TTX. (H-I) Sample traces confirming that the A23a→ aCC synapse is GABAergic. Cells were recorded, as previously described, before (black trace) and after (gray trace) bath application of 10 µM Picrotoxin (H) or 1 mM Gabazine (I), two blockers of the Drosophila GABA A receptor. In both cases, aCC inputs were abolished.

    Article Snippet: The CLI-Gal4 line, less specific compared to the other ones, exhibited the largest synaptic drive to aCC, which is notably reduced, but not fully blocked, by the presence of TTX (−9.55 ± 3.19 pA/pF, n = 4, to -3.57 ± 0.92 pA/pF, n = 6, ).

    Techniques: Activation Assay, Injection, Inhibition, Isolation

    ChR; NaChBac is a powerful tool to verify monosynaptic connectivity. (A) A representative current-clamp recording from the A27h interneuron overexpressing both ChR and NaChBac. Optogenetic stimulation (λ470 nm, 1 s) induced activation of NaChBac which persists in presence of 2 µM TTX (arrowheads). Conversely, APs produced by activation of endogenous voltage-gated sodium channels were blocked after TTX application (arrows). (B) Voltage dependence of NaChBac activation recorded from A27h in current-clamp. A27h depolarisation was elicited by injecting constant current steps (1 pA steps/0.5 s) in the presence of TTX. (C-D) Voltage-dependent inactivation of NaChBac. Peak amplitude was recorded and measured from A27h held at different voltages (from -90 to -20 mV) during optogenetic stimulation (λ470 nm, 1 s). NaChBac activation is affected at potentials more positive than -40 mV. Note: there is a second activation (peak) of NaChBaC at -90mV. (D) Averaged data ± SEM ( n = 3). (E) Sample recording of synaptic drive to aCC, recorded in voltage-clamp, following optogenetic activation of A27h (λ470 nm, 1 s). In presence of TTX, co-expression and activation of both ChR and NaChBac in A27h produced a clear synaptic input in aCC (inward current, black trace), thus confirming the existence of a monosynaptic connection between these two neurons. As a control, TTX successfully blocked aCC inputs when only ChR, but not NaChBac, was expressed in A27h (red trace).

    Journal: bioRxiv

    Article Title: Electrophysiological validation of premotor interneurons monosynaptically connected to the aCC motoneuron in the Drosophila larval CNS

    doi: 10.1101/2020.06.17.156430

    Figure Lengend Snippet: ChR; NaChBac is a powerful tool to verify monosynaptic connectivity. (A) A representative current-clamp recording from the A27h interneuron overexpressing both ChR and NaChBac. Optogenetic stimulation (λ470 nm, 1 s) induced activation of NaChBac which persists in presence of 2 µM TTX (arrowheads). Conversely, APs produced by activation of endogenous voltage-gated sodium channels were blocked after TTX application (arrows). (B) Voltage dependence of NaChBac activation recorded from A27h in current-clamp. A27h depolarisation was elicited by injecting constant current steps (1 pA steps/0.5 s) in the presence of TTX. (C-D) Voltage-dependent inactivation of NaChBac. Peak amplitude was recorded and measured from A27h held at different voltages (from -90 to -20 mV) during optogenetic stimulation (λ470 nm, 1 s). NaChBac activation is affected at potentials more positive than -40 mV. Note: there is a second activation (peak) of NaChBaC at -90mV. (D) Averaged data ± SEM ( n = 3). (E) Sample recording of synaptic drive to aCC, recorded in voltage-clamp, following optogenetic activation of A27h (λ470 nm, 1 s). In presence of TTX, co-expression and activation of both ChR and NaChBac in A27h produced a clear synaptic input in aCC (inward current, black trace), thus confirming the existence of a monosynaptic connection between these two neurons. As a control, TTX successfully blocked aCC inputs when only ChR, but not NaChBac, was expressed in A27h (red trace).

    Article Snippet: The CLI-Gal4 line, less specific compared to the other ones, exhibited the largest synaptic drive to aCC, which is notably reduced, but not fully blocked, by the presence of TTX (−9.55 ± 3.19 pA/pF, n = 4, to -3.57 ± 0.92 pA/pF, n = 6, ).

    Techniques: Activation Assay, Produced, Expressing

    Effects of barbaloin on ATX II-induced late sodium current ( I Na.L ) enhancements and under normal conditions in rabbit ventricular myocytes. (A) Representative single traces of the effects of 4 μmol/L TTX on ATX II-induced I Na.L enhancement. (B) Representative single traces showing that barbaloin exerted reversible inhibitory effects on ATX II-induced I Na.L enhancement. (C) Representative single traces of the effects of 200 μmol/L barbaloin on I Na.L under normal conditions. (D) Single traces of the effects of barbaloin on ATX II-induced I Na.L enhancements at a membrane potential of −20 mV. (E) Representative traces of the effects of barbaloin on ATX II-induced I Na.L enhancements at membrane potentials of −80, −60, −50, −40 and −20 mV. (F) Bar graphs showing the mean ATX II-increased I Na.L percentage values for the control, 100 μmol/L barbaloin-treated and wash-out groups ( n =10. ** P

    Journal: Acta Pharmacologica Sinica

    Article Title: Barbaloin inhibits ventricular arrhythmias in rabbits by modulating voltage-gated ion channels

    doi: 10.1038/aps.2017.93

    Figure Lengend Snippet: Effects of barbaloin on ATX II-induced late sodium current ( I Na.L ) enhancements and under normal conditions in rabbit ventricular myocytes. (A) Representative single traces of the effects of 4 μmol/L TTX on ATX II-induced I Na.L enhancement. (B) Representative single traces showing that barbaloin exerted reversible inhibitory effects on ATX II-induced I Na.L enhancement. (C) Representative single traces of the effects of 200 μmol/L barbaloin on I Na.L under normal conditions. (D) Single traces of the effects of barbaloin on ATX II-induced I Na.L enhancements at a membrane potential of −20 mV. (E) Representative traces of the effects of barbaloin on ATX II-induced I Na.L enhancements at membrane potentials of −80, −60, −50, −40 and −20 mV. (F) Bar graphs showing the mean ATX II-increased I Na.L percentage values for the control, 100 μmol/L barbaloin-treated and wash-out groups ( n =10. ** P

    Article Snippet: ATX II and TTX were purchased from the Alomone Labs (Jerusalem, Israel) and Tocris (Ellisville, MO, USA), respectively.

    Techniques: