retigabine  (Alomone Labs)


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    Name:
    Retigabine
    Description:
    Retigabine is a potent and selective Kv7 KCNQ M channel modulator enhancer which is used in the clinic to treat epilepsy Retigabine 0 1 to 10 muM induced a K current and hyperpolarized CHO cells expressing Kv7 2 3 cells as well as other channels in the following order Kv7 3 Kv7 2 3 Kv7 2 Kv7 4 Similar effects were seen with 10 muM retigabine in oocytes expressing the Kv7 2 3 heteromeric channel
    Catalog Number:
    R-100
    Price:
    47.0
    Category:
    Small Molecule
    Source:
    Synthetic
    Applications:
    0
    Purity:
    >99% (HPLC)
    Size:
    10 mg
    Format:
    Lyophilized/solid.
    Formula:
    C16H18FN3O2
    Molecular Weight:
    303.3
    Molecule Name:
    Ethyl N-[2-amino-4-[(4-fluorophenyl)methylamino]phenyl]carbamate.
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    Structured Review

    Alomone Labs retigabine
    Retigabine
    Retigabine is a potent and selective Kv7 KCNQ M channel modulator enhancer which is used in the clinic to treat epilepsy Retigabine 0 1 to 10 muM induced a K current and hyperpolarized CHO cells expressing Kv7 2 3 cells as well as other channels in the following order Kv7 3 Kv7 2 3 Kv7 2 Kv7 4 Similar effects were seen with 10 muM retigabine in oocytes expressing the Kv7 2 3 heteromeric channel
    https://www.bioz.com/result/retigabine/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    retigabine - by Bioz Stars, 2021-09
    94/100 stars

    Images

    1) Product Images from "Mechanisms of PKA-Dependent Potentiation of Kv7.5 Channel Activity in Human Airway Smooth Muscle Cells"

    Article Title: Mechanisms of PKA-Dependent Potentiation of Kv7.5 Channel Activity in Human Airway Smooth Muscle Cells

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19082223

    Protein kinase A (PKA)-dependent regulation of endogenous Kv7.5 currents in cultured HASMCs. ( A ) Representative current traces recorded in a single HASMC (Capacitance = 281 pF) before (i. control) and 5 min after addition of 10 µM retigabine (ii). ( B ) Mean fractional conductance plot calculated from steady-state endogenous Kv7 currents fitted to a Boltzmann distribution (V 0.5 = −40.8 mV, n = 10). ( C ) I–V relationships of Kv7 currents recorded in HASMCs before (control, filled circles, n = 5), after 5 min treatment with 1 µM forskolin (open circles, n = 4), and after 5 min treatment with diclofenac (100 µM, open triangles, n = 4). ( D ) I–V relationships of Kv7 currents recorded in HASMCs before (control, filled circles, n = 7), after 5 min treatment with 1 nM formoterol (open circles, n = 7), and after 5 min treatment with Kv7 channel blocker XE991 (1 µM) in the presence of 1 nM formoterol (closed triangles, n = 3). ( E ) Representative time-courses of 1 nM formoterol application recorded at −20 mV in a single untreated HASMC (black, Capacitance = 39 pF) and a HASMC pretreated with 10 µM H-89 (red, C = 127 pF). ( F ) Relative formoterol-induced enhancement of the current recorded at −20 mV in untreated HASMCs (black bars, n = 7) and in HASMCs pretreated with H-89 (10 µM for 20 min, red bar, n = 4). * Significant difference from control ( p
    Figure Legend Snippet: Protein kinase A (PKA)-dependent regulation of endogenous Kv7.5 currents in cultured HASMCs. ( A ) Representative current traces recorded in a single HASMC (Capacitance = 281 pF) before (i. control) and 5 min after addition of 10 µM retigabine (ii). ( B ) Mean fractional conductance plot calculated from steady-state endogenous Kv7 currents fitted to a Boltzmann distribution (V 0.5 = −40.8 mV, n = 10). ( C ) I–V relationships of Kv7 currents recorded in HASMCs before (control, filled circles, n = 5), after 5 min treatment with 1 µM forskolin (open circles, n = 4), and after 5 min treatment with diclofenac (100 µM, open triangles, n = 4). ( D ) I–V relationships of Kv7 currents recorded in HASMCs before (control, filled circles, n = 7), after 5 min treatment with 1 nM formoterol (open circles, n = 7), and after 5 min treatment with Kv7 channel blocker XE991 (1 µM) in the presence of 1 nM formoterol (closed triangles, n = 3). ( E ) Representative time-courses of 1 nM formoterol application recorded at −20 mV in a single untreated HASMC (black, Capacitance = 39 pF) and a HASMC pretreated with 10 µM H-89 (red, C = 127 pF). ( F ) Relative formoterol-induced enhancement of the current recorded at −20 mV in untreated HASMCs (black bars, n = 7) and in HASMCs pretreated with H-89 (10 µM for 20 min, red bar, n = 4). * Significant difference from control ( p

    Techniques Used: Cell Culture

    2) Product Images from "The KV7 channel activator retigabine suppresses mouse urinary bladder afferent nerve activity without affecting detrusor smooth muscle K+ channel currents"

    Article Title: The KV7 channel activator retigabine suppresses mouse urinary bladder afferent nerve activity without affecting detrusor smooth muscle K+ channel currents

    Journal: The Journal of Physiology

    doi: 10.1113/JP277021

    K V 7 channel modulators alter TCs and afferent activity A–D , representative traces of bladder pressure (top) and afferent nerve activity (mean frequency, bottom) during ex vivo bladder filling, before drug exposure ( A ) and after addition of the K V 7 channel activator retigabine (10 μM, B ), the K V 7 channel blocker XE‐991 (10 μM, C ), or XE‐991 and retigabine combined ( D ). While nearly abolished, discernable TCs still occurred in the presence of retigabine ( B , inset). E–G , summary bar graphs showing TC integral ( E ), TC frequency ( F ), and peak afferent nerve activity ( G ) in the absence or presence of retigabine, XE‐991, or a combination of both drugs. Retigabine (10 μM) significantly reduced TC amplitude and peak afferent activity without significantly reducing leading slope, effects that were blocked by XE‐991. XE‐991 alone had no effect on TC amplitude, TC slope or afferent activity, but did cause a significant increase in TC frequency that was reversed by retigabine. H , summary graph showing that baseline afferent nerve activity (black circles) is nearly abolished by retigabine at all pressures, but is significantly augmented by XE‐991 only at intravesical pressures ≥ 18 mmHg (open circles). Both effects were blocked by incubation with the combination of the two drugs. E–G , * P
    Figure Legend Snippet: K V 7 channel modulators alter TCs and afferent activity A–D , representative traces of bladder pressure (top) and afferent nerve activity (mean frequency, bottom) during ex vivo bladder filling, before drug exposure ( A ) and after addition of the K V 7 channel activator retigabine (10 μM, B ), the K V 7 channel blocker XE‐991 (10 μM, C ), or XE‐991 and retigabine combined ( D ). While nearly abolished, discernable TCs still occurred in the presence of retigabine ( B , inset). E–G , summary bar graphs showing TC integral ( E ), TC frequency ( F ), and peak afferent nerve activity ( G ) in the absence or presence of retigabine, XE‐991, or a combination of both drugs. Retigabine (10 μM) significantly reduced TC amplitude and peak afferent activity without significantly reducing leading slope, effects that were blocked by XE‐991. XE‐991 alone had no effect on TC amplitude, TC slope or afferent activity, but did cause a significant increase in TC frequency that was reversed by retigabine. H , summary graph showing that baseline afferent nerve activity (black circles) is nearly abolished by retigabine at all pressures, but is significantly augmented by XE‐991 only at intravesical pressures ≥ 18 mmHg (open circles). Both effects were blocked by incubation with the combination of the two drugs. E–G , * P

    Techniques Used: Activity Assay, Ex Vivo, Incubation

    Concentration dependent effects of retigabine and XE‐991 on TCs and associated afferent activity A–C , summary bar graphs showing TC integral ( A ), peak afferent nerve activity ( B ), and TC frequency ( C ) in the absence or presence of retigabine (10 nM to 10 μM). Retigabine reduced TC amplitude and associated afferent activity in a concentration‐dependent manner. D–F , summary bar graphs showing TC integral ( D ), peak afferent nerve activity ( E ), and TC frequency ( F ) in the absence or presence of XE‐991 (10 nM to 10 μM). XE‐991 did not significantly change reduced TC amplitude, TC leading slope, or afferent activity; TC frequency was only increased with 10 μM XE‐991. * P
    Figure Legend Snippet: Concentration dependent effects of retigabine and XE‐991 on TCs and associated afferent activity A–C , summary bar graphs showing TC integral ( A ), peak afferent nerve activity ( B ), and TC frequency ( C ) in the absence or presence of retigabine (10 nM to 10 μM). Retigabine reduced TC amplitude and associated afferent activity in a concentration‐dependent manner. D–F , summary bar graphs showing TC integral ( D ), peak afferent nerve activity ( E ), and TC frequency ( F ) in the absence or presence of XE‐991 (10 nM to 10 μM). XE‐991 did not significantly change reduced TC amplitude, TC leading slope, or afferent activity; TC frequency was only increased with 10 μM XE‐991. * P

    Techniques Used: Concentration Assay, Activity Assay

    Retigabine inhibits Ca 2+ currents in freshly isolated mouse UBSM cells A , representative trace of inward Ca 2+ currents in response to a 20 ms voltage step from −60 to +10 mV. B , retigabine (10 μM) significantly reduced inward Ca 2+ currents in isolated UBSM cells. C–E , isometric contractility of urinary bladder strips in response to 60 mM K + in the absence ( C ) or presence ( D ) of increasing concentrations of retigabine. Upward, square deflections in D are an artefact of tissue oxygenation and are not muscle contractions. Retigabine (1–10 μM) reduced steady‐state contractile responses to 60 mM K + as compared to vehicle (DMSO) ( E ). * P
    Figure Legend Snippet: Retigabine inhibits Ca 2+ currents in freshly isolated mouse UBSM cells A , representative trace of inward Ca 2+ currents in response to a 20 ms voltage step from −60 to +10 mV. B , retigabine (10 μM) significantly reduced inward Ca 2+ currents in isolated UBSM cells. C–E , isometric contractility of urinary bladder strips in response to 60 mM K + in the absence ( C ) or presence ( D ) of increasing concentrations of retigabine. Upward, square deflections in D are an artefact of tissue oxygenation and are not muscle contractions. Retigabine (1–10 μM) reduced steady‐state contractile responses to 60 mM K + as compared to vehicle (DMSO) ( E ). * P

    Techniques Used: Isolation, Mass Spectrometry

    Retigabine does not affect membrane potential in freshly isolated mouse UBSM cells A , representative trace of membrane potential of freshly isolated, current‐clamped UBSM cells in the presence of retigabine alone, retigabine with the K ATP channel opener pinacidil, and 140 mM extracellular K + . B , retigabine had no effect on membrane potential, whereas pinacidil caused hyperpolarization and 140 mM K + caused a reversible depolarization. Circles represent individual paired experiments. Bars represent mean ± SEM of each condition ( n = 3–7 cells from N = 3–5 mice).
    Figure Legend Snippet: Retigabine does not affect membrane potential in freshly isolated mouse UBSM cells A , representative trace of membrane potential of freshly isolated, current‐clamped UBSM cells in the presence of retigabine alone, retigabine with the K ATP channel opener pinacidil, and 140 mM extracellular K + . B , retigabine had no effect on membrane potential, whereas pinacidil caused hyperpolarization and 140 mM K + caused a reversible depolarization. Circles represent individual paired experiments. Bars represent mean ± SEM of each condition ( n = 3–7 cells from N = 3–5 mice).

    Techniques Used: Isolation, Mouse Assay

    Retigabine inhibits bursts of afferent activity during simulated TCs A–D , representative traces of intravesical pressure (top) and afferent nerve activity (bottom) during simulated TCs in the absence of drug ( A ) or in the presence of 10 μM retigabine ( B ), 10 μM XE‐991 ( C ), or both drugs ( D ). TCs were simulated by gentle compression of the bladder wall with plastic‐covered forceps. E , comparison of simulated TC peak afferent activity in the presence of retigabine, XE‐991, or a combination of both. Peak afferent activity per change in simulated TC integral was reduced by retigabine ( F ), an effect that was reversed by pre‐incubation of tissue with XE‐991. * P
    Figure Legend Snippet: Retigabine inhibits bursts of afferent activity during simulated TCs A–D , representative traces of intravesical pressure (top) and afferent nerve activity (bottom) during simulated TCs in the absence of drug ( A ) or in the presence of 10 μM retigabine ( B ), 10 μM XE‐991 ( C ), or both drugs ( D ). TCs were simulated by gentle compression of the bladder wall with plastic‐covered forceps. E , comparison of simulated TC peak afferent activity in the presence of retigabine, XE‐991, or a combination of both. Peak afferent activity per change in simulated TC integral was reduced by retigabine ( F ), an effect that was reversed by pre‐incubation of tissue with XE‐991. * P

    Techniques Used: Activity Assay, Incubation

    Retigabine inhibits all afferent nerve activity A–B , representative traces of intravesical pressure (top) and afferent nerve activity (bottom) in the presence of the L‐type Ca 2+ channel blocker diltiazem ( A ) or the K V 7 agonist retigabine ( B ). C , diltiazem (grey circles) blocked all transient contractions and associated bursts of nerve activity, but did not affect baseline afferent activity. Retigabine (black circles) not only abolished TCs and associated afferent activity, but also eliminated the remaining baseline afferent nerve activity. * P
    Figure Legend Snippet: Retigabine inhibits all afferent nerve activity A–B , representative traces of intravesical pressure (top) and afferent nerve activity (bottom) in the presence of the L‐type Ca 2+ channel blocker diltiazem ( A ) or the K V 7 agonist retigabine ( B ). C , diltiazem (grey circles) blocked all transient contractions and associated bursts of nerve activity, but did not affect baseline afferent activity. Retigabine (black circles) not only abolished TCs and associated afferent activity, but also eliminated the remaining baseline afferent nerve activity. * P

    Techniques Used: Activity Assay

    Retigabine‐sensitive inward currents are absent in the presence of 95 mM extracellular K + A , representative recordings of inward currents in response to the addition of 95 mM extracellular K + . B , summary graph showing that retigabine did not alter inward currents, whereas the K ATP channel opener pinacidil caused a significant inward K + current. Circles represent individual paired experiments. Bars represent mean ± SEM of each condition. * P
    Figure Legend Snippet: Retigabine‐sensitive inward currents are absent in the presence of 95 mM extracellular K + A , representative recordings of inward currents in response to the addition of 95 mM extracellular K + . B , summary graph showing that retigabine did not alter inward currents, whereas the K ATP channel opener pinacidil caused a significant inward K + current. Circles represent individual paired experiments. Bars represent mean ± SEM of each condition. * P

    Techniques Used:

    3) Product Images from "Modulation of KV7 Channel Deactivation by PI(4,5)P2"

    Article Title: Modulation of KV7 Channel Deactivation by PI(4,5)P2

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2020.00895

    To determine whether the decrease in the rate of deactivation was a unique property of the heteromeric K V 7.2/K V 7.3 channel. (A, B) K + -current recordings from oocytes expressing K V 7.2 in the absence and presence of 10μM Retigabine (RTG). (C) τ DEACT was plotted as a function of t PULSE for the homomeric K V 7.2. It was found that the slow down of the deactvation kinetics still occurs when K V 7.2 was expressed alone, suggesting that the hysteretic behavior of these K V 7 channels was not a property emerging from heteromerization ( n = 5). (D) τ DEACT -t PULSE plots in C, replotted with a logarithmic t PULSE to highlight the behavior of τ DEACT at short t PULSE values.
    Figure Legend Snippet: To determine whether the decrease in the rate of deactivation was a unique property of the heteromeric K V 7.2/K V 7.3 channel. (A, B) K + -current recordings from oocytes expressing K V 7.2 in the absence and presence of 10μM Retigabine (RTG). (C) τ DEACT was plotted as a function of t PULSE for the homomeric K V 7.2. It was found that the slow down of the deactvation kinetics still occurs when K V 7.2 was expressed alone, suggesting that the hysteretic behavior of these K V 7 channels was not a property emerging from heteromerization ( n = 5). (D) τ DEACT -t PULSE plots in C, replotted with a logarithmic t PULSE to highlight the behavior of τ DEACT at short t PULSE values.

    Techniques Used: Expressing

    PI(4,5)P 2 regulates the action of Retigabine. (A–C) K + -currents were activated with a +40 mV pulses of variable duration in Xenopus oocytes expression K V 7.2/K V 7.3 channels. Subsequently, the currents were deactivated at −90 mV. These recordings were performed in the absence (A) and the presence of 1 µM (blue diamonds, n = 6) and 5 µM of Retigabine (red triangles, n = 7) (C) . (D–F) A two-exponential function was fitted to deactivating currents and the yielded τ 1 , τ 2 and the fractional contribution of the second component (fraction of τ 2 ) were plotted against t PULSE . ( D–F , respectively). (G–I) Semi-logarithmic version of the plot in (D–F) , detailing τ DEACT for t PULSE up to 1 second. For reference, the equivalent values from Figure 2 were plotted as small black square and small red circles. Black arrows in panels (D–I) indicate the range of t PULSE values at which values were statistically different between the recordings in the presence of 1 µM and 5 µM Retigabine.
    Figure Legend Snippet: PI(4,5)P 2 regulates the action of Retigabine. (A–C) K + -currents were activated with a +40 mV pulses of variable duration in Xenopus oocytes expression K V 7.2/K V 7.3 channels. Subsequently, the currents were deactivated at −90 mV. These recordings were performed in the absence (A) and the presence of 1 µM (blue diamonds, n = 6) and 5 µM of Retigabine (red triangles, n = 7) (C) . (D–F) A two-exponential function was fitted to deactivating currents and the yielded τ 1 , τ 2 and the fractional contribution of the second component (fraction of τ 2 ) were plotted against t PULSE . ( D–F , respectively). (G–I) Semi-logarithmic version of the plot in (D–F) , detailing τ DEACT for t PULSE up to 1 second. For reference, the equivalent values from Figure 2 were plotted as small black square and small red circles. Black arrows in panels (D–I) indicate the range of t PULSE values at which values were statistically different between the recordings in the presence of 1 µM and 5 µM Retigabine.

    Techniques Used: Expressing

    Overall deactivation kinetics as a function of the t PULSE . (A) As before, to consolidate the analysis of the deactivation kinetics, τ DEACT was plotted against the duration of the +40-mV Pulse (t PULSE ) for currents recorded in the presence of 1 μM and 5 μM Retigabine (blue open diamonds and red oepn hexagons, respectively). (B) The values of τ DEACT were significantly different for t PULSE longer than 61 ms. For reference, the equivalent values from Figure 2 were plotted as black solid square and red solid circles. (C, D) Equivalent τ DEACT -t PULSE plots generated from Wortmannin-treated oocytes.
    Figure Legend Snippet: Overall deactivation kinetics as a function of the t PULSE . (A) As before, to consolidate the analysis of the deactivation kinetics, τ DEACT was plotted against the duration of the +40-mV Pulse (t PULSE ) for currents recorded in the presence of 1 μM and 5 μM Retigabine (blue open diamonds and red oepn hexagons, respectively). (B) The values of τ DEACT were significantly different for t PULSE longer than 61 ms. For reference, the equivalent values from Figure 2 were plotted as black solid square and red solid circles. (C, D) Equivalent τ DEACT -t PULSE plots generated from Wortmannin-treated oocytes.

    Techniques Used: Generated

    4) Product Images from "Removal of KCNQ2 from Parvalbumin-expressing Interneurons Improves Anti-Seizure Efficacy of Retigabine"

    Article Title: Removal of KCNQ2 from Parvalbumin-expressing Interneurons Improves Anti-Seizure Efficacy of Retigabine

    Journal: bioRxiv

    doi: 10.1101/2020.12.09.417295

    Effect of retigabine (RTG) on the passive membrane properties of hippocampal PV-INs in vitro . (A) Bath application of RTG in hippocampal slices significantly hyperpolarized the RMP of PV-INs from WT mice (n=20, ** p
    Figure Legend Snippet: Effect of retigabine (RTG) on the passive membrane properties of hippocampal PV-INs in vitro . (A) Bath application of RTG in hippocampal slices significantly hyperpolarized the RMP of PV-INs from WT mice (n=20, ** p

    Techniques Used: In Vitro, Mouse Assay

    Effects of retigabine (RTG) on the passive membrane properties of hippocampal CA1-PCs in vitro. (A) Genotype had a significant effect on baseline RMP of CA1-PCs (n=19 for WT, n=24 cKO, * p
    Figure Legend Snippet: Effects of retigabine (RTG) on the passive membrane properties of hippocampal CA1-PCs in vitro. (A) Genotype had a significant effect on baseline RMP of CA1-PCs (n=19 for WT, n=24 cKO, * p

    Techniques Used: In Vitro

    Effects of retigabine (RTG) on the active excitability of PV-INs in vitro. (A) Experimental configuration in CA1. Whole cell recordings were obtained simultaneously from four neurons, including PV-INs (1, 2) and PCs (3, 4) before and after bath application of RTG (10 µM). PV-INs were distinguished from PCs and other interneurons by firing pattern (above) and post hoc morphology recovery (below). (B) Representative membrane voltage responses to different current step injections in PV-INs from WT (left) and cKO (right) before and after RTG application. RTG suppressed APs in PV-INs in WT (C, n=21, ** p
    Figure Legend Snippet: Effects of retigabine (RTG) on the active excitability of PV-INs in vitro. (A) Experimental configuration in CA1. Whole cell recordings were obtained simultaneously from four neurons, including PV-INs (1, 2) and PCs (3, 4) before and after bath application of RTG (10 µM). PV-INs were distinguished from PCs and other interneurons by firing pattern (above) and post hoc morphology recovery (below). (B) Representative membrane voltage responses to different current step injections in PV-INs from WT (left) and cKO (right) before and after RTG application. RTG suppressed APs in PV-INs in WT (C, n=21, ** p

    Techniques Used: In Vitro

    5) Product Images from "Reversal of a treatment-resistant, depression-related brain state with the Kv7 channel opener retigabine"

    Article Title: Reversal of a treatment-resistant, depression-related brain state with the Kv7 channel opener retigabine

    Journal: Neuroscience

    doi: 10.1016/j.neuroscience.2019.03.003

    Retigabine reduces I h currents and normalizes neural excitability of HFD-exposed animals to control levels. A ) Current injection data from layer II/III PLC pyramidal cells of HFD-exposed mice before and after bath application of 100 μM retigabine, with representative traces shown on the left and summary data on the right. We observed significant main effects of current injected (F (20,160) = 23.07, p
    Figure Legend Snippet: Retigabine reduces I h currents and normalizes neural excitability of HFD-exposed animals to control levels. A ) Current injection data from layer II/III PLC pyramidal cells of HFD-exposed mice before and after bath application of 100 μM retigabine, with representative traces shown on the left and summary data on the right. We observed significant main effects of current injected (F (20,160) = 23.07, p

    Techniques Used: Injection, Planar Chromatography, Mouse Assay

    6) Product Images from ""

    Article Title:

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    doi: 10.1124/jpet.117.241679

    Retigabine shifted effective potentials of XE991 to more negative potentials. (A) Voltage protocol and representative traces showing that when cells were pretreated with 10 µ M retigabine, 25-second treatment with 10 µ M XE991 (black box) inhibited Kv7.2 current at a holding potential of −70 mV. (B) Voltage-XE991 inhibition relationships and activation curve for Kv7.2 channels pretreated with 10 µ M retigabine. (C) Activation-XE991 inhibition relationships for Kv7.2 channel from results shown in (B). Error bars show S.E.M.
    Figure Legend Snippet: Retigabine shifted effective potentials of XE991 to more negative potentials. (A) Voltage protocol and representative traces showing that when cells were pretreated with 10 µ M retigabine, 25-second treatment with 10 µ M XE991 (black box) inhibited Kv7.2 current at a holding potential of −70 mV. (B) Voltage-XE991 inhibition relationships and activation curve for Kv7.2 channels pretreated with 10 µ M retigabine. (C) Activation-XE991 inhibition relationships for Kv7.2 channel from results shown in (B). Error bars show S.E.M.

    Techniques Used: Inhibition, Activation Assay

    Kv7.2 mutant (R214D) shifted voltage dependence of activation and XE991 inhibition to more positive potentials. (A) Voltage protocol and representative traces showing that 25-second treatment with 10 µ M XE991 (black box) inhibited WT Kv7.2 current at a holding potential of −40 mV, whereas Kv7.2(R214D) mutant channel had minimal inhibition at this holding potential. (B) Voltage-XE991 inhibition relationships and activation curve for Kv7.2(R214D) channels. (C) Activation–XE991 inhibition relationships for Kv7.2(R214D) channel from results shown in (B). (D) Pooled results for V 1/2 inhibition and V 1/2 activation potentials of Kv7.2, retigabine-treated Kv7.2 and Kv7.2(R214D) channel. Slope of regression line is also shown. Error bars show S.E.M.
    Figure Legend Snippet: Kv7.2 mutant (R214D) shifted voltage dependence of activation and XE991 inhibition to more positive potentials. (A) Voltage protocol and representative traces showing that 25-second treatment with 10 µ M XE991 (black box) inhibited WT Kv7.2 current at a holding potential of −40 mV, whereas Kv7.2(R214D) mutant channel had minimal inhibition at this holding potential. (B) Voltage-XE991 inhibition relationships and activation curve for Kv7.2(R214D) channels. (C) Activation–XE991 inhibition relationships for Kv7.2(R214D) channel from results shown in (B). (D) Pooled results for V 1/2 inhibition and V 1/2 activation potentials of Kv7.2, retigabine-treated Kv7.2 and Kv7.2(R214D) channel. Slope of regression line is also shown. Error bars show S.E.M.

    Techniques Used: Mutagenesis, Activation Assay, Inhibition

    7) Product Images from "The anticonvulsant retigabine is a subtype selective modulator of GABA receptors"

    Article Title: The anticonvulsant retigabine is a subtype selective modulator of GABA receptors

    Journal: Epilepsia

    doi: 10.1111/epi.12950

    Effects of retigabine on GABA ‐evoked currents in tsA 201 cells expressing various combinations of GABA A receptor subunits. Currents through α1β2 ( A ), α4β3 ( B ), α6β2 ( C ), α1β2γ2S ( D ), α4β3γ2S ( E ), α6β2γ2S ( F ), α1β2δ ( G ), α4β3δ ( H ), and α6β2δ ( I ) receptors were evoked by the application of the indicated concentrations of GABA in the presence of solvent (control; black) or 10 μ m retigabine (blue). For original sample traces see Figure 4 A,C. For the concentration–response curves, all peak current amplitudes determined in one cell were normalized to the amplitude of the current triggered by 300 μ m GABA in the presence of solvent. Data were fitted to a Hill equation; parameters are given in Table 1 .
    Figure Legend Snippet: Effects of retigabine on GABA ‐evoked currents in tsA 201 cells expressing various combinations of GABA A receptor subunits. Currents through α1β2 ( A ), α4β3 ( B ), α6β2 ( C ), α1β2γ2S ( D ), α4β3γ2S ( E ), α6β2γ2S ( F ), α1β2δ ( G ), α4β3δ ( H ), and α6β2δ ( I ) receptors were evoked by the application of the indicated concentrations of GABA in the presence of solvent (control; black) or 10 μ m retigabine (blue). For original sample traces see Figure 4 A,C. For the concentration–response curves, all peak current amplitudes determined in one cell were normalized to the amplitude of the current triggered by 300 μ m GABA in the presence of solvent. Data were fitted to a Hill equation; parameters are given in Table 1 .

    Techniques Used: Expressing, Concentration Assay

    Retigabine concentration–response relation for GABA ‐evoked currents through α1β2 or α1β2δ GABA A receptors expressed in tsA 201 cells. Currents were evoked by the application of either 2 μ m ( A and B ) or 6 μ m ( C and D ) GABA (the EC 50 values for α1β2 and α1β2δ receptors, respectively; see Table 1 ), in the presence of either solvent or the indicated concentrations of retigabine (ret). ( A , C ) Original traces of currents evoked by GABA in a cell expressing either α1β2 ( A ) or α1β2δ ( C ). ( B , D ) Concentration–response curves for the effect of retigabine on GABA ‐induced currents in cells expressing either α1β2 ( B ; orange) or α1β2δ ( D ; red). Peak current amplitudes determined in the presence of the indicated concentrations of retigabine were normalized to the amplitudes obtained in the presence of solvent (n = 8). *, ***, **** indicate significant differences versus the amplitudes obtained in the presence of solvent at p
    Figure Legend Snippet: Retigabine concentration–response relation for GABA ‐evoked currents through α1β2 or α1β2δ GABA A receptors expressed in tsA 201 cells. Currents were evoked by the application of either 2 μ m ( A and B ) or 6 μ m ( C and D ) GABA (the EC 50 values for α1β2 and α1β2δ receptors, respectively; see Table 1 ), in the presence of either solvent or the indicated concentrations of retigabine (ret). ( A , C ) Original traces of currents evoked by GABA in a cell expressing either α1β2 ( A ) or α1β2δ ( C ). ( B , D ) Concentration–response curves for the effect of retigabine on GABA ‐induced currents in cells expressing either α1β2 ( B ; orange) or α1β2δ ( D ; red). Peak current amplitudes determined in the presence of the indicated concentrations of retigabine were normalized to the amplitudes obtained in the presence of solvent (n = 8). *, ***, **** indicate significant differences versus the amplitudes obtained in the presence of solvent at p

    Techniques Used: Concentration Assay, Expressing

    Effects of retigabine on seizure‐like activity and tonic currents in cultured hippocampal neurons. Seizure‐like activity was induced by low extracellular Mg 2+ (Mg 2+ ‐free solution), XE 991, or bicuculline (bic) applied either alone or together. ( A – D ) Original recordings of seizure‐like activity induced by low Mg 2+ ( A ), 30 μ m XE 991 ( B ), 30 μ m bicuculline (bic; C ), or XE 991 plus bicuculline ( D ). Traces were obtained before (left) and during (right) the presence of 3 μ m retigabine (ret). Scale bars: 1 s. Dashed lines refer to the average membrane potential prior to the induction of seizure‐like activity. ( E ) Concentration–response curve for the inhibition of seizure‐like activity induced by low Mg 2+ (orange) in the presence of the indicated concentrations of retigabine. This inhibition was evaluated by the reduction of the area under the curve ( AUC ), which was half maximal at 1.3 ± 0.3 μ m (n = 7). The inhibitory effect of 3 μ m retigabine was also quantified for seizure‐like activity induced by XE 991 (red), bicuculline (bic; green), or XE 991 plus bicuculline (green‐blue). AUC values obtained with XE 991 plus bicuculline were significantly different from all other values at p
    Figure Legend Snippet: Effects of retigabine on seizure‐like activity and tonic currents in cultured hippocampal neurons. Seizure‐like activity was induced by low extracellular Mg 2+ (Mg 2+ ‐free solution), XE 991, or bicuculline (bic) applied either alone or together. ( A – D ) Original recordings of seizure‐like activity induced by low Mg 2+ ( A ), 30 μ m XE 991 ( B ), 30 μ m bicuculline (bic; C ), or XE 991 plus bicuculline ( D ). Traces were obtained before (left) and during (right) the presence of 3 μ m retigabine (ret). Scale bars: 1 s. Dashed lines refer to the average membrane potential prior to the induction of seizure‐like activity. ( E ) Concentration–response curve for the inhibition of seizure‐like activity induced by low Mg 2+ (orange) in the presence of the indicated concentrations of retigabine. This inhibition was evaluated by the reduction of the area under the curve ( AUC ), which was half maximal at 1.3 ± 0.3 μ m (n = 7). The inhibitory effect of 3 μ m retigabine was also quantified for seizure‐like activity induced by XE 991 (red), bicuculline (bic; green), or XE 991 plus bicuculline (green‐blue). AUC values obtained with XE 991 plus bicuculline were significantly different from all other values at p

    Techniques Used: Activity Assay, Cell Culture, Concentration Assay, Inhibition

    Effects of retigabine on GABA ‐evoked currents in cultured hippocampal neurons. Currents were evoked by the application of the indicated concentrations of GABA in either solvent (control) or 5 m m penicillin, 10 μ m retigabine, or both. ( A ) Original traces of currents evoked by the indicated concentrations of GABA in one hippocampal neuron in the presence of solvent (control; black) or 10 μ m retigabine (blue). ( B ) Concentration–response curves for GABA ‐evoked currents in the presence of solvent (control; black) or 10 μ m retigabine (blue; n = 7). All peak current amplitudes determined in one neuron were normalized to the amplitude of the current triggered by 100 μ m GABA in the presence of solvent. ( C ) Original traces of currents evoked by the indicated concentrations of GABA in one hippocampal neuron in the presence of solvent (control: black) or 5 m m penicillin (red). ( D ) Concentration response curves for GABA ‐evoked currents in the presence of solvent (control; black) or 5 m m penicillin (red; n = 5). All peak current amplitudes determined in one neuron were normalized to the amplitude of the current triggered by 100 μ m GABA in the presence of solvent. ( E ) Original traces of currents were evoked by the indicated concentrations of GABA in one hippocampal neuron in the presence of either 5 m m penicillin (red) or 5 m m penicillin plus 10 μ m retigabine (blue). ( F ) Concentration–response curves for GABA ‐evoked currents in the presence of either 5 m m penicillin (red) or 5 m m penicillin plus 10 μ m retigabine (blue; n = 7). All peak current amplitudes determined in one neuron were normalized to the amplitude of the current triggered by 100 μ m GABA in the presence of penicillin only. Maximal GABA current amplitudes were significantly larger in the presence of retigabine (p
    Figure Legend Snippet: Effects of retigabine on GABA ‐evoked currents in cultured hippocampal neurons. Currents were evoked by the application of the indicated concentrations of GABA in either solvent (control) or 5 m m penicillin, 10 μ m retigabine, or both. ( A ) Original traces of currents evoked by the indicated concentrations of GABA in one hippocampal neuron in the presence of solvent (control; black) or 10 μ m retigabine (blue). ( B ) Concentration–response curves for GABA ‐evoked currents in the presence of solvent (control; black) or 10 μ m retigabine (blue; n = 7). All peak current amplitudes determined in one neuron were normalized to the amplitude of the current triggered by 100 μ m GABA in the presence of solvent. ( C ) Original traces of currents evoked by the indicated concentrations of GABA in one hippocampal neuron in the presence of solvent (control: black) or 5 m m penicillin (red). ( D ) Concentration response curves for GABA ‐evoked currents in the presence of solvent (control; black) or 5 m m penicillin (red; n = 5). All peak current amplitudes determined in one neuron were normalized to the amplitude of the current triggered by 100 μ m GABA in the presence of solvent. ( E ) Original traces of currents were evoked by the indicated concentrations of GABA in one hippocampal neuron in the presence of either 5 m m penicillin (red) or 5 m m penicillin plus 10 μ m retigabine (blue). ( F ) Concentration–response curves for GABA ‐evoked currents in the presence of either 5 m m penicillin (red) or 5 m m penicillin plus 10 μ m retigabine (blue; n = 7). All peak current amplitudes determined in one neuron were normalized to the amplitude of the current triggered by 100 μ m GABA in the presence of penicillin only. Maximal GABA current amplitudes were significantly larger in the presence of retigabine (p

    Techniques Used: Cell Culture, Concentration Assay

    8) Product Images from "Differential Potassium Channel Gene Regulation in BXD Mice Reveals Novel Targets for Pharmacogenetic Therapies to Reduce Heavy Alcohol Drinking"

    Article Title: Differential Potassium Channel Gene Regulation in BXD Mice Reveals Novel Targets for Pharmacogenetic Therapies to Reduce Heavy Alcohol Drinking

    Journal: Alcohol (Fayetteville, N.Y.)

    doi: 10.1016/j.alcohol.2016.05.007

    The Kv7 channel positive modulator, retigabine, reduced ethanol consumption in high-drinking, but not low-drinking male C57BL/6J mice ( A ) Average weekly ethanol consumption and ( B ) intra-strain variability of 24 hr ethanol consumption in C57BL/6J mice during week 7 of the intermittent access model. ( C ) Despite the large between-subjects variation, the coefficient of variance during 7 weeks of access to ethanol in the intermittent model indicates increased within-subject stability (n = 19 mice, * p
    Figure Legend Snippet: The Kv7 channel positive modulator, retigabine, reduced ethanol consumption in high-drinking, but not low-drinking male C57BL/6J mice ( A ) Average weekly ethanol consumption and ( B ) intra-strain variability of 24 hr ethanol consumption in C57BL/6J mice during week 7 of the intermittent access model. ( C ) Despite the large between-subjects variation, the coefficient of variance during 7 weeks of access to ethanol in the intermittent model indicates increased within-subject stability (n = 19 mice, * p

    Techniques Used: Mouse Assay

    9) Product Images from "Hyper-SUMOylation of the Kv7 Potassium Channel Diminishes the M-Current Leading to Seizures and Sudden Death"

    Article Title: Hyper-SUMOylation of the Kv7 Potassium Channel Diminishes the M-Current Leading to Seizures and Sudden Death

    Journal: Neuron

    doi: 10.1016/j.neuron.2014.07.042

    Prevention of Acoustics-Induced Seizures and AV Block in SENP2 fxN/fxN Mice with Retigabine
    Figure Legend Snippet: Prevention of Acoustics-Induced Seizures and AV Block in SENP2 fxN/fxN Mice with Retigabine

    Techniques Used: Blocking Assay, Mouse Assay

    10) Product Images from "The anticonvulsant retigabine suppresses neuronal KV2-mediated currents"

    Article Title: The anticonvulsant retigabine suppresses neuronal KV2-mediated currents

    Journal: Scientific Reports

    doi: 10.1038/srep35080

    Retigabine inhibition of K V 2.1 is voltage-dependent. ( a ) Typical current recordings of K V 2.1 channels to determine the activation (left) and inactivation (right) properties, before (top) and after exposure to 100 μM retigabine (bottom). Voltage protocols are shown on top. ( b ) Concentration-effect relationship of K V 2.1 inhibition. Retigabine inhibition of K V 2.1 (closed circles) currents was not significantly different (p = 0.385) in the presence of KCNE2 (open circles). ( c ) Voltage-dependence of activation (circles) and inactivation (triangles) in absence (closed symbols) and presence (open symbols) of 100 μM retigabine. The voltage-dependence of activation was obtained by plotting the normalized tail currents (I/I max ) in the activation current traces from panel A as function of the prepulse potential. Retigabine induced a small but significant (p = 0.012) hyperpolarizing shift in the voltage-dependence of activation. The voltage-dependence of inactivation, obtained by plotting the normalized peak current (I/I max ) at +60 mV after a 5 s prepulse as a function of the prepulse potential, was not affected by retigabine. ( d ) Time constants of K V 2.1 channel opening (≥0 mV) and closing (
    Figure Legend Snippet: Retigabine inhibition of K V 2.1 is voltage-dependent. ( a ) Typical current recordings of K V 2.1 channels to determine the activation (left) and inactivation (right) properties, before (top) and after exposure to 100 μM retigabine (bottom). Voltage protocols are shown on top. ( b ) Concentration-effect relationship of K V 2.1 inhibition. Retigabine inhibition of K V 2.1 (closed circles) currents was not significantly different (p = 0.385) in the presence of KCNE2 (open circles). ( c ) Voltage-dependence of activation (circles) and inactivation (triangles) in absence (closed symbols) and presence (open symbols) of 100 μM retigabine. The voltage-dependence of activation was obtained by plotting the normalized tail currents (I/I max ) in the activation current traces from panel A as function of the prepulse potential. Retigabine induced a small but significant (p = 0.012) hyperpolarizing shift in the voltage-dependence of activation. The voltage-dependence of inactivation, obtained by plotting the normalized peak current (I/I max ) at +60 mV after a 5 s prepulse as a function of the prepulse potential, was not affected by retigabine. ( d ) Time constants of K V 2.1 channel opening (≥0 mV) and closing (

    Techniques Used: Inhibition, Activation Assay, Concentration Assay

    Retigabine inhibits most K V channels in the intermediate to high μM range. A two-step pulse protocol adjusted to the biophysical properties of the respective channel was used. The voltage applied is shown below the respective K V channel current traces. Retigabine (colored traces) inhibited all K V channels in the high μM range ( > 100 μM) with exception of K V 2.1, which was inhibited at relative low μM concentrations (10 μM).
    Figure Legend Snippet: Retigabine inhibits most K V channels in the intermediate to high μM range. A two-step pulse protocol adjusted to the biophysical properties of the respective channel was used. The voltage applied is shown below the respective K V channel current traces. Retigabine (colored traces) inhibited all K V channels in the high μM range ( > 100 μM) with exception of K V 2.1, which was inhibited at relative low μM concentrations (10 μM).

    Techniques Used:

    Retigabine inhibits the K V 2-mediated component of the outward current in cultured rat hippocampal neurons. ( a ) Representative current traces from cultured rat hippocampal neurons. 100 μM retigabine inhibited the outward current and the RTG-sensitive current was obtained after subtraction. 100 nM Guangxitoxin-1E (GxTx), i.e. selective K V 2 inhibitor, was used to confirm the inhibition of K V 2-mediated current by retigabine. Retigabine inhibited a major component of delayed rectifier current, with little inhibition caused by GxTx ( b ) similar to ( a ) although the K V 2-mediated current was first inhibited with GxTx before applying retigabine. Inhibition of the K V 2-mediated component of the current by GxTx resulted in little inhibition of retigabine. However, retigabine still inhibited a fast activating and inactivating current. ( c,d ) Current-voltage relationship, obtained by plotting the total outward current at the end of the 250 ms step as function of the voltage with retigabine ( c ) or GxTx ( d ) initial exposure. ( e,f ) Fractional inhibition as a function of the applied voltage. As observed in HEK cells ( Fig. 3f ), retigabine ( e ) had a voltage-dependence of inhibition that could be abolished after subsequent exposure to GxTx. Panel ( f ) is similar to ( e ) but with initial exposure to GxTx. Lines represent the voltage-dependence of activation fitted with the Boltzmann equation.
    Figure Legend Snippet: Retigabine inhibits the K V 2-mediated component of the outward current in cultured rat hippocampal neurons. ( a ) Representative current traces from cultured rat hippocampal neurons. 100 μM retigabine inhibited the outward current and the RTG-sensitive current was obtained after subtraction. 100 nM Guangxitoxin-1E (GxTx), i.e. selective K V 2 inhibitor, was used to confirm the inhibition of K V 2-mediated current by retigabine. Retigabine inhibited a major component of delayed rectifier current, with little inhibition caused by GxTx ( b ) similar to ( a ) although the K V 2-mediated current was first inhibited with GxTx before applying retigabine. Inhibition of the K V 2-mediated component of the current by GxTx resulted in little inhibition of retigabine. However, retigabine still inhibited a fast activating and inactivating current. ( c,d ) Current-voltage relationship, obtained by plotting the total outward current at the end of the 250 ms step as function of the voltage with retigabine ( c ) or GxTx ( d ) initial exposure. ( e,f ) Fractional inhibition as a function of the applied voltage. As observed in HEK cells ( Fig. 3f ), retigabine ( e ) had a voltage-dependence of inhibition that could be abolished after subsequent exposure to GxTx. Panel ( f ) is similar to ( e ) but with initial exposure to GxTx. Lines represent the voltage-dependence of activation fitted with the Boltzmann equation.

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

    Inhibition of K V 2.1 current by retigabine is only partly reversible. ( a ) Representative K V 2.1 current traces (black) at +30 mV (left). The scaled current traces are shown in the right panel. Retigabine (grey) inhibited approximately 80% of the current but inhibition was poorly recovered 30 minutes after removal of retigabine (dotted). The ‘apparent’ acceleration of the inactivation process seen in the scaled current traces most likely reflects open-channel block by retigabine. ( b ) Plot of a representative wash-in/wash-out experiment. Inhibition of the K V 2.1 current occurred slowly, typically requiring 5–10 minutes to achieve saturation. Inhibition of K V 2.1 currents was poorly reversible and occurred extremely slow. ( c ) Bar chart illustrating the degree of current (I/I c ), with I the current at a given condition and I c the control condition. K V 2.1 inhibition was poorly reversible, independent of the solvent, and significantly different from current rundown. *Indicates statistical significance (p
    Figure Legend Snippet: Inhibition of K V 2.1 current by retigabine is only partly reversible. ( a ) Representative K V 2.1 current traces (black) at +30 mV (left). The scaled current traces are shown in the right panel. Retigabine (grey) inhibited approximately 80% of the current but inhibition was poorly recovered 30 minutes after removal of retigabine (dotted). The ‘apparent’ acceleration of the inactivation process seen in the scaled current traces most likely reflects open-channel block by retigabine. ( b ) Plot of a representative wash-in/wash-out experiment. Inhibition of the K V 2.1 current occurred slowly, typically requiring 5–10 minutes to achieve saturation. Inhibition of K V 2.1 currents was poorly reversible and occurred extremely slow. ( c ) Bar chart illustrating the degree of current (I/I c ), with I the current at a given condition and I c the control condition. K V 2.1 inhibition was poorly reversible, independent of the solvent, and significantly different from current rundown. *Indicates statistical significance (p

    Techniques Used: Inhibition, Blocking Assay

    Molecular pharmacology on K V 2 and K V 7 channels in HEK cells compared with retigabine plasma concentrations. ( a ) Concentration-effect relationship of retigabine potentiation on K V 7.2-K V 7.3 currents in the absence (filled circles) and presence (open circles) of KCNE2. The grey bar represents the plasma concentration range, minimum (0.65 μM) to maximum (6.6 μM), in patients treated with 600–1200 mg retigabine/day 38 39 40 41 42 . Although KCNE2 shifted the concentration-effect curve, K V 7.2-K V 7.3 current potentiation was not fully prevented in the plasma concentration range. ( b ) Concentration-effect relationship of K V 2.1 inhibition in absence (blue, filled circles) and presence (blue, open circles) of KCNE2, obtained from direct perfusion of retigabine on the Kv2.1 channels. The light grey bar represents the plasma concentration range as in ( a ). Black, white, dark grey and striped bar represent the x-fold reduction in K V 2.1 current density obtained from the retigabine incubation experiments. Although little direct K V 2.1 inhibition occurred, maximal suppression of the K V 2.1 current density occurred in the plasma concentration range.
    Figure Legend Snippet: Molecular pharmacology on K V 2 and K V 7 channels in HEK cells compared with retigabine plasma concentrations. ( a ) Concentration-effect relationship of retigabine potentiation on K V 7.2-K V 7.3 currents in the absence (filled circles) and presence (open circles) of KCNE2. The grey bar represents the plasma concentration range, minimum (0.65 μM) to maximum (6.6 μM), in patients treated with 600–1200 mg retigabine/day 38 39 40 41 42 . Although KCNE2 shifted the concentration-effect curve, K V 7.2-K V 7.3 current potentiation was not fully prevented in the plasma concentration range. ( b ) Concentration-effect relationship of K V 2.1 inhibition in absence (blue, filled circles) and presence (blue, open circles) of KCNE2, obtained from direct perfusion of retigabine on the Kv2.1 channels. The light grey bar represents the plasma concentration range as in ( a ). Black, white, dark grey and striped bar represent the x-fold reduction in K V 2.1 current density obtained from the retigabine incubation experiments. Although little direct K V 2.1 inhibition occurred, maximal suppression of the K V 2.1 current density occurred in the plasma concentration range.

    Techniques Used: Concentration Assay, Inhibition, Incubation

    KCNE2 decreases the retigabine sensitivity of heterotetrameric K V 7.2-K V 7.3 channels. ( a ) Effect of increasing concentrations of retigabine (1–100 μM) on K V 7.2-K V 7.3 currents. Retigabine potentiated the K V 7.2-K V 7.3 current in a concentration-dependent manner, and saturation occurred above 30 μM. Voltage protocol is shown on top. ( b ) Voltage-dependence of activation. Increasing concentrations of RTG caused a gradual hyperpolarizing shift. ( c ) Concentration-effect curve plotted as the shift in the voltage-dependence of activation normalized to the maximal observed shift (ΔV/ΔV max ) as function of the drug concentration. ( d ) Similar to ( a ) but after co-expression with KCNE2. Retigabine potentiated the K V 7.2-K V 7.3-KCNE2 currents but unlike ( a ) concentrations above 1 μM had to be used. ( e ) Voltage-dependence of activation. KCNE2 reduced the hyperpolarizing shift at every drug concentration, and decreased the maximal observed shift (ΔV max ). ( f ) Concentration-effect curve.
    Figure Legend Snippet: KCNE2 decreases the retigabine sensitivity of heterotetrameric K V 7.2-K V 7.3 channels. ( a ) Effect of increasing concentrations of retigabine (1–100 μM) on K V 7.2-K V 7.3 currents. Retigabine potentiated the K V 7.2-K V 7.3 current in a concentration-dependent manner, and saturation occurred above 30 μM. Voltage protocol is shown on top. ( b ) Voltage-dependence of activation. Increasing concentrations of RTG caused a gradual hyperpolarizing shift. ( c ) Concentration-effect curve plotted as the shift in the voltage-dependence of activation normalized to the maximal observed shift (ΔV/ΔV max ) as function of the drug concentration. ( d ) Similar to ( a ) but after co-expression with KCNE2. Retigabine potentiated the K V 7.2-K V 7.3-KCNE2 currents but unlike ( a ) concentrations above 1 μM had to be used. ( e ) Voltage-dependence of activation. KCNE2 reduced the hyperpolarizing shift at every drug concentration, and decreased the maximal observed shift (ΔV max ). ( f ) Concentration-effect curve.

    Techniques Used: Concentration Assay, Activation Assay, Expressing

    11) Product Images from "Retigabine holds KV7 channels open and stabilizes the resting potential"

    Article Title: Retigabine holds KV7 channels open and stabilizes the resting potential

    Journal: The Journal of General Physiology

    doi: 10.1085/jgp.201511517

    Effect of Retigabine on the activity of the heteromeric K V 7.2/K V 7.3 channel. (A) K + currents recorded in the absence or presence of 1–5-µM Retigabine using the same protocol shown in Fig. 1 . (B) Average normalized K + current amplitude measured at the beginning of the deactivating currents (I TAIL ; arrows in A). Normalized I TAIL versus test pulse potential (I TAIL –V PULSE ) plots incrementally shift toward negative potentials as the concentration of Retigabine (RTG) was increased. Individual I TAIL –V PULSE plots were fitted to a double Boltzmann distribution, and the weighted average half-maximum potential (weighted V 1/2 ) was plotted as a function of the concentration of Retigabine (inset). Although the weighted V 1/2 values were not statistically different, this parameter changed to more negative potentials as a function of the concentration of Retigabine in all our recordings. Error bars represent standard deviation. n = 5–8.
    Figure Legend Snippet: Effect of Retigabine on the activity of the heteromeric K V 7.2/K V 7.3 channel. (A) K + currents recorded in the absence or presence of 1–5-µM Retigabine using the same protocol shown in Fig. 1 . (B) Average normalized K + current amplitude measured at the beginning of the deactivating currents (I TAIL ; arrows in A). Normalized I TAIL versus test pulse potential (I TAIL –V PULSE ) plots incrementally shift toward negative potentials as the concentration of Retigabine (RTG) was increased. Individual I TAIL –V PULSE plots were fitted to a double Boltzmann distribution, and the weighted average half-maximum potential (weighted V 1/2 ) was plotted as a function of the concentration of Retigabine (inset). Although the weighted V 1/2 values were not statistically different, this parameter changed to more negative potentials as a function of the concentration of Retigabine in all our recordings. Error bars represent standard deviation. n = 5–8.

    Techniques Used: Activity Assay, Concentration Assay, Standard Deviation

    The resting potential, but not the threshold potential for triggering AP, was affected by Retigabine. (A) APs were recorded from Xenopus oocytes coexpressing both the α and β subunits of Na V 1.4, ShakerIR, K V 7.2, and K V 7.3. The APs were recorded using the loose two-electrode voltage-clamp technique (see Materials and methods for details) in the absence (black trace) or presence of 1-µM (red trace) or 10-µM (green trace) Retigabine. The gray trace shows the response of V M when a subthreshold was applied. The arrow indicates the moment at which AP threshold (V THR ) is reached. (B) Effect of Retigabine (RTG) on the resting membrane potential (V REST ) and AP threshold (V THR ). (C) Minimum current injection needed to evoke an AP (I THR ). The values of I THR were normalized by I THR in the absence of Retigabine ( n = 4). (D) Example of a phase plot calculated from the APs shown on A. V REST was strongly affected by Retigabine (blue star), whereas V THR remained fairly unaltered. Although the depolarization phase was affected by Retigabine (blue arrow), the late phase of repolarization (yellow arrow) remained unaltered. Arrows indicate the progress of the AP in time during the stimulation phase (purple arrow), depolarization phase (green arrow), and repolarization phase (orange arrow). The open blue arrow points at the times when APs reached their maximum voltage. The open yellow arrow points at the late repolarization phase. Error bars represent standard deviation. t test; n = 5. *, P
    Figure Legend Snippet: The resting potential, but not the threshold potential for triggering AP, was affected by Retigabine. (A) APs were recorded from Xenopus oocytes coexpressing both the α and β subunits of Na V 1.4, ShakerIR, K V 7.2, and K V 7.3. The APs were recorded using the loose two-electrode voltage-clamp technique (see Materials and methods for details) in the absence (black trace) or presence of 1-µM (red trace) or 10-µM (green trace) Retigabine. The gray trace shows the response of V M when a subthreshold was applied. The arrow indicates the moment at which AP threshold (V THR ) is reached. (B) Effect of Retigabine (RTG) on the resting membrane potential (V REST ) and AP threshold (V THR ). (C) Minimum current injection needed to evoke an AP (I THR ). The values of I THR were normalized by I THR in the absence of Retigabine ( n = 4). (D) Example of a phase plot calculated from the APs shown on A. V REST was strongly affected by Retigabine (blue star), whereas V THR remained fairly unaltered. Although the depolarization phase was affected by Retigabine (blue arrow), the late phase of repolarization (yellow arrow) remained unaltered. Arrows indicate the progress of the AP in time during the stimulation phase (purple arrow), depolarization phase (green arrow), and repolarization phase (orange arrow). The open blue arrow points at the times when APs reached their maximum voltage. The open yellow arrow points at the late repolarization phase. Error bars represent standard deviation. t test; n = 5. *, P

    Techniques Used: Injection, Standard Deviation

    Deactivation time constant increase as a function of the duration of the activating pulse. (A and B) K + currents were activated by 40-mV pulses with a duration varying from 10 to 4,175 ms in the absence (A) and presence (B) of 1-µM Retigabine. The duration of the pulse increased 1.3-fold for each trace. After activation, the K + current was deactivated at −90 mV. (C) τ DEACT –t PULSE plot shows that τ DEACT increased as the activating pulse was longer (black squares). In the presence of Retigabine, τ DEACT increased further. (D) The same plot as in C, but the t PULSE is displayed in a logarithmic scale to highlight the τ DEACT –t PULSE relationship for shorter activating pulses and showing that τ DEACT was unaltered by Retigabine for pulses shorter than 500 ms. Error bars represent standard deviation. Control, n = 7; Retigabine, n = 6.
    Figure Legend Snippet: Deactivation time constant increase as a function of the duration of the activating pulse. (A and B) K + currents were activated by 40-mV pulses with a duration varying from 10 to 4,175 ms in the absence (A) and presence (B) of 1-µM Retigabine. The duration of the pulse increased 1.3-fold for each trace. After activation, the K + current was deactivated at −90 mV. (C) τ DEACT –t PULSE plot shows that τ DEACT increased as the activating pulse was longer (black squares). In the presence of Retigabine, τ DEACT increased further. (D) The same plot as in C, but the t PULSE is displayed in a logarithmic scale to highlight the τ DEACT –t PULSE relationship for shorter activating pulses and showing that τ DEACT was unaltered by Retigabine for pulses shorter than 500 ms. Error bars represent standard deviation. Control, n = 7; Retigabine, n = 6.

    Techniques Used: Activation Assay, Standard Deviation

    Deactivation from the depolarized and resting potential open states. (A) Deactivation at −90 mV of the K + current elicited by a 100-ms depolarization to 40 mV in the absence (black trace) and presence of 1-µM Retigabine (red trace). (B) Deactivation at −90 mV of the K + current elicited by holding the membrane potential at −50 mV also in the absence (black trace) and presence of 1-µM Retigabine (red trace). (C–-F) Deactivation time constant (τ DEACT ) versus deactivating potential (τ DEACT –V DEACT ) plot of channels activated by a 100-ms depolarization to 40 mV (C) or by holding the membrane potential at −60 mV, −50 mV, and −40 mV (D, E, and F, respectively) in the absence (black squares) and presence of 1-µM Retigabine (RTG; red circles). Error bars represent standard deviation. t test; n = 5–8. *, P
    Figure Legend Snippet: Deactivation from the depolarized and resting potential open states. (A) Deactivation at −90 mV of the K + current elicited by a 100-ms depolarization to 40 mV in the absence (black trace) and presence of 1-µM Retigabine (red trace). (B) Deactivation at −90 mV of the K + current elicited by holding the membrane potential at −50 mV also in the absence (black trace) and presence of 1-µM Retigabine (red trace). (C–-F) Deactivation time constant (τ DEACT ) versus deactivating potential (τ DEACT –V DEACT ) plot of channels activated by a 100-ms depolarization to 40 mV (C) or by holding the membrane potential at −60 mV, −50 mV, and −40 mV (D, E, and F, respectively) in the absence (black squares) and presence of 1-µM Retigabine (RTG; red circles). Error bars represent standard deviation. t test; n = 5–8. *, P

    Techniques Used: Standard Deviation

    Proposed kinetic scheme for the activity of K V 7.2/K V 7.3 and the effect of Retigabine. The scheme contains five global states, which are different with respect to each other depending on the activation status of the VSD and the pore domain (P). In the scheme, VSD represents the four voltage sensors of the channel. At negative potentials, the VSDs are in the deactivated state (VSD D ) and the pore is, therefore, closed (P C0 ). Upon depolarization, the VSDs are activated (VSD A ), allowing the pore to sojourn between a conductive and a nonconductive state (P O1 and P C1 , respectively). Holding the membrane depolarized potentials keeps the VSDs in the VSD A state while allowing a second transition in the pore. As a result, the pore sojourns between a second pair of conductive and nonconductive states (P O2 and P C2 , respectively) and the channel becomes Retigabine sensitive. Deactivation from these states is slower than from the pair P C1 –P O1 . Retigabine further decreases the rate of deactivation from the pair P C2 –P O2 while leaving deactivation unaffected from the other pair.
    Figure Legend Snippet: Proposed kinetic scheme for the activity of K V 7.2/K V 7.3 and the effect of Retigabine. The scheme contains five global states, which are different with respect to each other depending on the activation status of the VSD and the pore domain (P). In the scheme, VSD represents the four voltage sensors of the channel. At negative potentials, the VSDs are in the deactivated state (VSD D ) and the pore is, therefore, closed (P C0 ). Upon depolarization, the VSDs are activated (VSD A ), allowing the pore to sojourn between a conductive and a nonconductive state (P O1 and P C1 , respectively). Holding the membrane depolarized potentials keeps the VSDs in the VSD A state while allowing a second transition in the pore. As a result, the pore sojourns between a second pair of conductive and nonconductive states (P O2 and P C2 , respectively) and the channel becomes Retigabine sensitive. Deactivation from these states is slower than from the pair P C1 –P O1 . Retigabine further decreases the rate of deactivation from the pair P C2 –P O2 while leaving deactivation unaffected from the other pair.

    Techniques Used: Activity Assay, Activation Assay

    12) Product Images from "Kv7 voltage-activated potassium channel inhibitors reduce fluid resuscitation requirements after hemorrhagic shock in rats"

    Article Title: Kv7 voltage-activated potassium channel inhibitors reduce fluid resuscitation requirements after hemorrhagic shock in rats

    Journal: Journal of Biomedical Science

    doi: 10.1186/s12929-017-0316-1

    Effects of Kv7 channel modulators on blood pressure in normal rats. a . Intravenous injection of increasing doses of linopridine (0.1-6 mg/kg in 0.5 mL normal saline). Arrows indicate time points of drug injection. Open squares : Systolic blood pressure. Grey squares : Diastolic blood pressure. Black Squares : Mean arterial blood pressure. BP: Blood pressure (mmHg). N = 3. Data are mean ± SD. b . Intravenous injection of increasing doses of retigabine (0.1-12 mg/kg in 0.5 mL normal saline) followed by an intravenous injection of 6 mg/kg linopirdine in 0.5 mL normal saline. Open squares : Systolic blood pressure. Grey squares : Diastolic blood pressure. Black Squares : Mean arterial blood pressure. BP: Blood pressure (mmHg). N = 3
    Figure Legend Snippet: Effects of Kv7 channel modulators on blood pressure in normal rats. a . Intravenous injection of increasing doses of linopridine (0.1-6 mg/kg in 0.5 mL normal saline). Arrows indicate time points of drug injection. Open squares : Systolic blood pressure. Grey squares : Diastolic blood pressure. Black Squares : Mean arterial blood pressure. BP: Blood pressure (mmHg). N = 3. Data are mean ± SD. b . Intravenous injection of increasing doses of retigabine (0.1-12 mg/kg in 0.5 mL normal saline) followed by an intravenous injection of 6 mg/kg linopirdine in 0.5 mL normal saline. Open squares : Systolic blood pressure. Grey squares : Diastolic blood pressure. Black Squares : Mean arterial blood pressure. BP: Blood pressure (mmHg). N = 3

    Techniques Used: Injection

    13) Product Images from "Differential Potassium Channel Gene Regulation in BXD Mice Reveals Novel Targets for Pharmacogenetic Therapies to Reduce Heavy Alcohol Drinking"

    Article Title: Differential Potassium Channel Gene Regulation in BXD Mice Reveals Novel Targets for Pharmacogenetic Therapies to Reduce Heavy Alcohol Drinking

    Journal: Alcohol (Fayetteville, N.Y.)

    doi: 10.1016/j.alcohol.2016.05.007

    The Kv7 channel positive modulator, retigabine, reduced ethanol consumption in high-drinking, but not low-drinking male C57BL/6J mice ( A ) Average weekly ethanol consumption and ( B ) intra-strain variability of 24 hr ethanol consumption in C57BL/6J mice during week 7 of the intermittent access model. ( C ) Despite the large between-subjects variation, the coefficient of variance during 7 weeks of access to ethanol in the intermittent model indicates increased within-subject stability (n = 19 mice, * p
    Figure Legend Snippet: The Kv7 channel positive modulator, retigabine, reduced ethanol consumption in high-drinking, but not low-drinking male C57BL/6J mice ( A ) Average weekly ethanol consumption and ( B ) intra-strain variability of 24 hr ethanol consumption in C57BL/6J mice during week 7 of the intermittent access model. ( C ) Despite the large between-subjects variation, the coefficient of variance during 7 weeks of access to ethanol in the intermittent model indicates increased within-subject stability (n = 19 mice, * p

    Techniques Used: Mouse Assay

    14) Product Images from "Effect of M-current modulation on mammalian vestibular responses to transient head motion"

    Article Title: Effect of M-current modulation on mammalian vestibular responses to transient head motion

    Journal: Journal of Neurophysiology

    doi: 10.1152/jn.00384.2017

    Effects of retigabine on vestibular responses. A : changes in vestibular response waveforms. Responses shown were recorded before (Bas, baseline) and 20 and 120 minutes after a retigabine dose of 20 mg/kg. After 20 min, responses were delayed (P1, N1, P2) and reduced in size (P1-N1, P2-N1). Thereafter, responses were enhanced, increasing in amplitude, decreasing in latency, and lowering of threshold (120 min). Stimulus level was 6 dB re: 1 g /ms. Calibration bars indicate 2.0 ms and 1.0 μ/V for graphic. Dashed vertical lines identify latencies of P1 and P2 before drug injection. B : effects of a single dose of retigabine (filled circles) and DMSO vehicle alone (open circles) on VsEP responses. Mean values are normalized to baseline (mean value minus mean baseline) for response latencies (P1, N1), amplitudes (P1-N1), and thresholds over a period of up to 2 h following retigabine (20 mg/kg; n = 6) and DMSO ( n = 9) injections. Data are means ± SD (retigabine group: black circles ± solid red lines; DMSO group: open circles ± blue dotted lines). Data to the right of the vertical dashed line represent the effects of drug or vehicle administration on VsEP responses. Baseline means were as follows: P1, 1.306 ± 0.033 ms; N1, 1.605 ± 0.071 ms; and P1-N1, 1.045 ± 0.246 μV; threshold = −10.5 ± 0.0 dB re: 1 g /ms. Stimulus level was 6 dB re: 1 g /ms.
    Figure Legend Snippet: Effects of retigabine on vestibular responses. A : changes in vestibular response waveforms. Responses shown were recorded before (Bas, baseline) and 20 and 120 minutes after a retigabine dose of 20 mg/kg. After 20 min, responses were delayed (P1, N1, P2) and reduced in size (P1-N1, P2-N1). Thereafter, responses were enhanced, increasing in amplitude, decreasing in latency, and lowering of threshold (120 min). Stimulus level was 6 dB re: 1 g /ms. Calibration bars indicate 2.0 ms and 1.0 μ/V for graphic. Dashed vertical lines identify latencies of P1 and P2 before drug injection. B : effects of a single dose of retigabine (filled circles) and DMSO vehicle alone (open circles) on VsEP responses. Mean values are normalized to baseline (mean value minus mean baseline) for response latencies (P1, N1), amplitudes (P1-N1), and thresholds over a period of up to 2 h following retigabine (20 mg/kg; n = 6) and DMSO ( n = 9) injections. Data are means ± SD (retigabine group: black circles ± solid red lines; DMSO group: open circles ± blue dotted lines). Data to the right of the vertical dashed line represent the effects of drug or vehicle administration on VsEP responses. Baseline means were as follows: P1, 1.306 ± 0.033 ms; N1, 1.605 ± 0.071 ms; and P1-N1, 1.045 ± 0.246 μV; threshold = −10.5 ± 0.0 dB re: 1 g /ms. Stimulus level was 6 dB re: 1 g /ms.

    Techniques Used: Mass Spectrometry, Injection

    15) Product Images from ""

    Article Title:

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    doi: 10.1124/jpet.117.241679

    Retigabine shifted effective potentials of XE991 to more negative potentials. (A) Voltage protocol and representative traces showing that when cells were pretreated with 10 µ M retigabine, 25-second treatment with 10 µ M XE991 (black box) inhibited Kv7.2 current at a holding potential of −70 mV. (B) Voltage-XE991 inhibition relationships and activation curve for Kv7.2 channels pretreated with 10 µ M retigabine. (C) Activation-XE991 inhibition relationships for Kv7.2 channel from results shown in (B). Error bars show S.E.M.
    Figure Legend Snippet: Retigabine shifted effective potentials of XE991 to more negative potentials. (A) Voltage protocol and representative traces showing that when cells were pretreated with 10 µ M retigabine, 25-second treatment with 10 µ M XE991 (black box) inhibited Kv7.2 current at a holding potential of −70 mV. (B) Voltage-XE991 inhibition relationships and activation curve for Kv7.2 channels pretreated with 10 µ M retigabine. (C) Activation-XE991 inhibition relationships for Kv7.2 channel from results shown in (B). Error bars show S.E.M.

    Techniques Used: Inhibition, Activation Assay

    Kv7.2 mutant (R214D) shifted voltage dependence of activation and XE991 inhibition to more positive potentials. (A) Voltage protocol and representative traces showing that 25-second treatment with 10 µ M XE991 (black box) inhibited WT Kv7.2 current at a holding potential of −40 mV, whereas Kv7.2(R214D) mutant channel had minimal inhibition at this holding potential. (B) Voltage-XE991 inhibition relationships and activation curve for Kv7.2(R214D) channels. (C) Activation–XE991 inhibition relationships for Kv7.2(R214D) channel from results shown in (B). (D) Pooled results for V 1/2 inhibition and V 1/2 activation potentials of Kv7.2, retigabine-treated Kv7.2 and Kv7.2(R214D) channel. Slope of regression line is also shown. Error bars show S.E.M.
    Figure Legend Snippet: Kv7.2 mutant (R214D) shifted voltage dependence of activation and XE991 inhibition to more positive potentials. (A) Voltage protocol and representative traces showing that 25-second treatment with 10 µ M XE991 (black box) inhibited WT Kv7.2 current at a holding potential of −40 mV, whereas Kv7.2(R214D) mutant channel had minimal inhibition at this holding potential. (B) Voltage-XE991 inhibition relationships and activation curve for Kv7.2(R214D) channels. (C) Activation–XE991 inhibition relationships for Kv7.2(R214D) channel from results shown in (B). (D) Pooled results for V 1/2 inhibition and V 1/2 activation potentials of Kv7.2, retigabine-treated Kv7.2 and Kv7.2(R214D) channel. Slope of regression line is also shown. Error bars show S.E.M.

    Techniques Used: Mutagenesis, Activation Assay, Inhibition

    16) Product Images from "Oxaliplatin Depolarizes the IB4– Dorsal Root Ganglion Neurons to Drive the Development of Neuropathic Pain Through TRPM8 in Mice"

    Article Title: Oxaliplatin Depolarizes the IB4– Dorsal Root Ganglion Neurons to Drive the Development of Neuropathic Pain Through TRPM8 in Mice

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2021.690858

    Effects of retigabine on the sensory and motor behaviors following oxaliplatin treatment. Oxaliplatin (oxa, 5 mg/kg, ip) was administrated at Day 0. Retigabine (10 mg/kg, ip, daily) was treated at Day –2, Day –1, Day 0, Day 1, and Day 2. The same groups of animals were tested for mechanical and cold behaviors at Day 3, and were tested for heat and motor behaviors at Day 4. (A) Paw withdrawal threshold to Von-Frey filament; (B) Score of withdrawal response to acetone; (C) Paw withdrawal latency to radiant heat; and (D) Time stayed on rod at different speeds in rotarod tests ( n = 6 for all groups). *, P
    Figure Legend Snippet: Effects of retigabine on the sensory and motor behaviors following oxaliplatin treatment. Oxaliplatin (oxa, 5 mg/kg, ip) was administrated at Day 0. Retigabine (10 mg/kg, ip, daily) was treated at Day –2, Day –1, Day 0, Day 1, and Day 2. The same groups of animals were tested for mechanical and cold behaviors at Day 3, and were tested for heat and motor behaviors at Day 4. (A) Paw withdrawal threshold to Von-Frey filament; (B) Score of withdrawal response to acetone; (C) Paw withdrawal latency to radiant heat; and (D) Time stayed on rod at different speeds in rotarod tests ( n = 6 for all groups). *, P

    Techniques Used:

    Effects of TC-I on the sensory and motor behaviors following oxaliplatin treatment. The administration of oxaliplatin, the schedule for TC-I (10 mg/kg, ip) treatment (same to the Retigabine treatment), the testing schedule for different types of behaviors were described in the Legend of Figure 3 . (A) Paw withdrawal threshold to Von-Frey filament; (B) Score of withdrawal response to acetone; (C) Paw withdrawal latency to radiant heat; and (D) Time stayed on rod at different speeds in rotarod tests ( n = 6 for all groups). **, P
    Figure Legend Snippet: Effects of TC-I on the sensory and motor behaviors following oxaliplatin treatment. The administration of oxaliplatin, the schedule for TC-I (10 mg/kg, ip) treatment (same to the Retigabine treatment), the testing schedule for different types of behaviors were described in the Legend of Figure 3 . (A) Paw withdrawal threshold to Von-Frey filament; (B) Score of withdrawal response to acetone; (C) Paw withdrawal latency to radiant heat; and (D) Time stayed on rod at different speeds in rotarod tests ( n = 6 for all groups). **, P

    Techniques Used:

    Effects of A-967079 on the sensory and motor behaviors following oxaliplatin treatment. The administration of oxaliplatin, the schedule for A-967079 (A-96, 100 mg/kg, po) treatment (same to the Retigabine and TC-I treatment), the testing schedule for different types of behaviors were described in the Legend of Figure 3 . (A) Paw withdrawal threshold to Von-Frey filament; (B) Score of withdrawal response to acetone; (C) Paw withdrawal latency to radiant heat; and (D) Time stayed on rod at different speeds in rotarod tests ( n = 6 for all groups).
    Figure Legend Snippet: Effects of A-967079 on the sensory and motor behaviors following oxaliplatin treatment. The administration of oxaliplatin, the schedule for A-967079 (A-96, 100 mg/kg, po) treatment (same to the Retigabine and TC-I treatment), the testing schedule for different types of behaviors were described in the Legend of Figure 3 . (A) Paw withdrawal threshold to Von-Frey filament; (B) Score of withdrawal response to acetone; (C) Paw withdrawal latency to radiant heat; and (D) Time stayed on rod at different speeds in rotarod tests ( n = 6 for all groups).

    Techniques Used:

    Related Articles

    Injection:

    Article Title: Nociceptor Translational Profiling Reveals the Ragulator-Rag GTPase Complex as a Critical Generator of Neuropathic Pain
    Article Snippet: .. Each morning, 0.1 ml of vehicle (saline) was injected intraperitoneally 10 min before confinement in the black chamber for 30 min. Three hours later, the same mouse was injected with retigabine (Alomone Labs) (10 mg/kg in 0.1 ml of saline). ..

    Article Title: CD3+ T cells are critical for the resolution of comorbid inflammatory pain and depression-like behavior
    Article Snippet: .. In the morning, mice were injected i.p. with phosphate buffered saline (PBS) and were individually placed 10 min later in the dark chamber for 15 min. Four hours later, mice were injected i.p. with 10 mg/kg of the analgesic drug retigabine i.p. (#R-100, Alomone laboratory, Jerusalem, Israel) and placed in the bright chamber. ..

    Mouse Assay:

    Article Title: CD3+ T cells are critical for the resolution of comorbid inflammatory pain and depression-like behavior
    Article Snippet: .. In the morning, mice were injected i.p. with phosphate buffered saline (PBS) and were individually placed 10 min later in the dark chamber for 15 min. Four hours later, mice were injected i.p. with 10 mg/kg of the analgesic drug retigabine i.p. (#R-100, Alomone laboratory, Jerusalem, Israel) and placed in the bright chamber. ..

    other:

    Article Title: The KV7 channel activator retigabine suppresses mouse urinary bladder afferent nerve activity without affecting detrusor smooth muscle K+ channel currents
    Article Snippet: Retigabine was obtained from Alomone Labs (Jerusalem, Israel).

    Article Title: Removal of KCNQ2 from Parvalbumin-expressing Interneurons Improves Anti-Seizure Efficacy of Retigabine
    Article Snippet: DrugsKainic Acid (KA; Santa Cruz Biotechnology) was dissolved in phosphate-buffered saline (PBS; Gibco, Life Technologies), and retigabine (RTG; Alomone Labs, Inc) and picrotoxin (PTX; Tocris) were first dissolved in dimethyl sulfoxide (DMSO) prior to being diluted to 1% volume/weight in PBS.

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    Alomone Labs retigabine
    Protein kinase A (PKA)-dependent regulation of endogenous Kv7.5 currents in cultured HASMCs. ( A ) Representative current traces recorded in a single HASMC (Capacitance = 281 pF) before (i. control) and 5 min after addition of 10 µM <t>retigabine</t> (ii). ( B ) Mean fractional conductance plot calculated from steady-state endogenous Kv7 currents fitted to a Boltzmann distribution (V 0.5 = −40.8 mV, n = 10). ( C ) I–V relationships of Kv7 currents recorded in HASMCs before (control, filled circles, n = 5), after 5 min treatment with 1 µM forskolin (open circles, n = 4), and after 5 min treatment with diclofenac (100 µM, open triangles, n = 4). ( D ) I–V relationships of Kv7 currents recorded in HASMCs before (control, filled circles, n = 7), after 5 min treatment with 1 nM formoterol (open circles, n = 7), and after 5 min treatment with Kv7 channel blocker XE991 (1 µM) in the presence of 1 nM formoterol (closed triangles, n = 3). ( E ) Representative time-courses of 1 nM formoterol application recorded at −20 mV in a single untreated HASMC (black, Capacitance = 39 pF) and a HASMC pretreated with 10 µM H-89 (red, C = 127 pF). ( F ) Relative formoterol-induced enhancement of the current recorded at −20 mV in untreated HASMCs (black bars, n = 7) and in HASMCs pretreated with H-89 (10 µM for 20 min, red bar, n = 4). * Significant difference from control ( p
    Retigabine, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Alomone Labs etb r
    HNa diet, in vivo: effects of an <t>ETB-R</t> antagonist on the ARNA responses to reflex increases in ERSNA.
    Etb R, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs ryanodine
    NT-3 generated long-range Ca 2+ signaling from the axon to the cell body. a Long-range Ca 2+ wave. The relative change in the Cal-520 emission ratio (defined as R) was used as a measure of changes in Ca 2+ concentration. The pseudocolored images represent R after local application of PBS ( top ) or NT-3 ( bottom ) to the axon ( arrow ). Scale bars, 50 μm. b The mean amplitude of R treat / R 0 for 90 s during local application of NT-3 in the presence of the indicated inhibitors (PBS = 10, NT-3 = 16, xestospongin C = 9, <t>ryanodine</t> = 15, dantrolene = 18, SKF96365 = 12 neurons from three independent experiments). c , d The axon was exposed to NT-3 in the presence of the indicated inhibitors, and then minor neurite outgrowth (PBS = 21, NT-3 = 21, xestospongin C = 26, ryanodine = 25, dantrolene = 25, SKF96365 = 24 neurites from three independent experiments) c and axonal outgrowth (PBS = 7, NT-3 = 9, xestospongin C = 9, ryanodine = 9, dantrolene = 8, SKF96365 = 8 neurons from three independent experiments) d were measured. e Local application of NT-3 increased the quantity of phospho-CaMKI in the cell body. After local application of PBS ( top ) or NT-3 ( bottom ), hippocampal neurons were immunostained with antibodies against CaMKI ( green ) and phospho-Thr177 of CaMKI ( magenta ). The merged images ( right panels ) are shown. The graph plots the fluorescence intensities of total CaMKI ( green ) and CaMKI phosphorylated at Thr177 ( magenta ) and in the line. Scale bars, 20 μm. f NT-3-induced minor neurite retraction was abolished by Ca 2+ signaling inhibitors. The axon was exposed to NT-3 in the presence of the indicated inhibitors, and minor neurite outgrowth was measured (PBS = 27, NT-3 = 31, BAPTA = 47, STO-609 = 45, KN-93 = 40 neurites from three independent experiments). g , h Local application of indicated inhibitors to the axon. Minor neurite outgrowth (DMSO = 42, xestospongin C = 32, ryanodine = 37, dantrolene = 35, SKF96365 = 32 neurons from three independent experiments) g and axonal outgrowth (DMSO = 14, xestospongin C = 11, ryanodine = 13, dantrolene = 13, SKF96365 = 12 neurons from three independent experiments) h were measured. Error bars represent SEM. * P
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    Protein kinase A (PKA)-dependent regulation of endogenous Kv7.5 currents in cultured HASMCs. ( A ) Representative current traces recorded in a single HASMC (Capacitance = 281 pF) before (i. control) and 5 min after addition of 10 µM retigabine (ii). ( B ) Mean fractional conductance plot calculated from steady-state endogenous Kv7 currents fitted to a Boltzmann distribution (V 0.5 = −40.8 mV, n = 10). ( C ) I–V relationships of Kv7 currents recorded in HASMCs before (control, filled circles, n = 5), after 5 min treatment with 1 µM forskolin (open circles, n = 4), and after 5 min treatment with diclofenac (100 µM, open triangles, n = 4). ( D ) I–V relationships of Kv7 currents recorded in HASMCs before (control, filled circles, n = 7), after 5 min treatment with 1 nM formoterol (open circles, n = 7), and after 5 min treatment with Kv7 channel blocker XE991 (1 µM) in the presence of 1 nM formoterol (closed triangles, n = 3). ( E ) Representative time-courses of 1 nM formoterol application recorded at −20 mV in a single untreated HASMC (black, Capacitance = 39 pF) and a HASMC pretreated with 10 µM H-89 (red, C = 127 pF). ( F ) Relative formoterol-induced enhancement of the current recorded at −20 mV in untreated HASMCs (black bars, n = 7) and in HASMCs pretreated with H-89 (10 µM for 20 min, red bar, n = 4). * Significant difference from control ( p

    Journal: International Journal of Molecular Sciences

    Article Title: Mechanisms of PKA-Dependent Potentiation of Kv7.5 Channel Activity in Human Airway Smooth Muscle Cells

    doi: 10.3390/ijms19082223

    Figure Lengend Snippet: Protein kinase A (PKA)-dependent regulation of endogenous Kv7.5 currents in cultured HASMCs. ( A ) Representative current traces recorded in a single HASMC (Capacitance = 281 pF) before (i. control) and 5 min after addition of 10 µM retigabine (ii). ( B ) Mean fractional conductance plot calculated from steady-state endogenous Kv7 currents fitted to a Boltzmann distribution (V 0.5 = −40.8 mV, n = 10). ( C ) I–V relationships of Kv7 currents recorded in HASMCs before (control, filled circles, n = 5), after 5 min treatment with 1 µM forskolin (open circles, n = 4), and after 5 min treatment with diclofenac (100 µM, open triangles, n = 4). ( D ) I–V relationships of Kv7 currents recorded in HASMCs before (control, filled circles, n = 7), after 5 min treatment with 1 nM formoterol (open circles, n = 7), and after 5 min treatment with Kv7 channel blocker XE991 (1 µM) in the presence of 1 nM formoterol (closed triangles, n = 3). ( E ) Representative time-courses of 1 nM formoterol application recorded at −20 mV in a single untreated HASMC (black, Capacitance = 39 pF) and a HASMC pretreated with 10 µM H-89 (red, C = 127 pF). ( F ) Relative formoterol-induced enhancement of the current recorded at −20 mV in untreated HASMCs (black bars, n = 7) and in HASMCs pretreated with H-89 (10 µM for 20 min, red bar, n = 4). * Significant difference from control ( p

    Article Snippet: Retigabine was from Alomone Labs (Jerusalem, Israel).

    Techniques: Cell Culture

    K V 7 channel modulators alter TCs and afferent activity A–D , representative traces of bladder pressure (top) and afferent nerve activity (mean frequency, bottom) during ex vivo bladder filling, before drug exposure ( A ) and after addition of the K V 7 channel activator retigabine (10 μM, B ), the K V 7 channel blocker XE‐991 (10 μM, C ), or XE‐991 and retigabine combined ( D ). While nearly abolished, discernable TCs still occurred in the presence of retigabine ( B , inset). E–G , summary bar graphs showing TC integral ( E ), TC frequency ( F ), and peak afferent nerve activity ( G ) in the absence or presence of retigabine, XE‐991, or a combination of both drugs. Retigabine (10 μM) significantly reduced TC amplitude and peak afferent activity without significantly reducing leading slope, effects that were blocked by XE‐991. XE‐991 alone had no effect on TC amplitude, TC slope or afferent activity, but did cause a significant increase in TC frequency that was reversed by retigabine. H , summary graph showing that baseline afferent nerve activity (black circles) is nearly abolished by retigabine at all pressures, but is significantly augmented by XE‐991 only at intravesical pressures ≥ 18 mmHg (open circles). Both effects were blocked by incubation with the combination of the two drugs. E–G , * P

    Journal: The Journal of Physiology

    Article Title: The KV7 channel activator retigabine suppresses mouse urinary bladder afferent nerve activity without affecting detrusor smooth muscle K+ channel currents

    doi: 10.1113/JP277021

    Figure Lengend Snippet: K V 7 channel modulators alter TCs and afferent activity A–D , representative traces of bladder pressure (top) and afferent nerve activity (mean frequency, bottom) during ex vivo bladder filling, before drug exposure ( A ) and after addition of the K V 7 channel activator retigabine (10 μM, B ), the K V 7 channel blocker XE‐991 (10 μM, C ), or XE‐991 and retigabine combined ( D ). While nearly abolished, discernable TCs still occurred in the presence of retigabine ( B , inset). E–G , summary bar graphs showing TC integral ( E ), TC frequency ( F ), and peak afferent nerve activity ( G ) in the absence or presence of retigabine, XE‐991, or a combination of both drugs. Retigabine (10 μM) significantly reduced TC amplitude and peak afferent activity without significantly reducing leading slope, effects that were blocked by XE‐991. XE‐991 alone had no effect on TC amplitude, TC slope or afferent activity, but did cause a significant increase in TC frequency that was reversed by retigabine. H , summary graph showing that baseline afferent nerve activity (black circles) is nearly abolished by retigabine at all pressures, but is significantly augmented by XE‐991 only at intravesical pressures ≥ 18 mmHg (open circles). Both effects were blocked by incubation with the combination of the two drugs. E–G , * P

    Article Snippet: Retigabine was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Activity Assay, Ex Vivo, Incubation

    Concentration dependent effects of retigabine and XE‐991 on TCs and associated afferent activity A–C , summary bar graphs showing TC integral ( A ), peak afferent nerve activity ( B ), and TC frequency ( C ) in the absence or presence of retigabine (10 nM to 10 μM). Retigabine reduced TC amplitude and associated afferent activity in a concentration‐dependent manner. D–F , summary bar graphs showing TC integral ( D ), peak afferent nerve activity ( E ), and TC frequency ( F ) in the absence or presence of XE‐991 (10 nM to 10 μM). XE‐991 did not significantly change reduced TC amplitude, TC leading slope, or afferent activity; TC frequency was only increased with 10 μM XE‐991. * P

    Journal: The Journal of Physiology

    Article Title: The KV7 channel activator retigabine suppresses mouse urinary bladder afferent nerve activity without affecting detrusor smooth muscle K+ channel currents

    doi: 10.1113/JP277021

    Figure Lengend Snippet: Concentration dependent effects of retigabine and XE‐991 on TCs and associated afferent activity A–C , summary bar graphs showing TC integral ( A ), peak afferent nerve activity ( B ), and TC frequency ( C ) in the absence or presence of retigabine (10 nM to 10 μM). Retigabine reduced TC amplitude and associated afferent activity in a concentration‐dependent manner. D–F , summary bar graphs showing TC integral ( D ), peak afferent nerve activity ( E ), and TC frequency ( F ) in the absence or presence of XE‐991 (10 nM to 10 μM). XE‐991 did not significantly change reduced TC amplitude, TC leading slope, or afferent activity; TC frequency was only increased with 10 μM XE‐991. * P

    Article Snippet: Retigabine was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Concentration Assay, Activity Assay

    Retigabine inhibits Ca 2+ currents in freshly isolated mouse UBSM cells A , representative trace of inward Ca 2+ currents in response to a 20 ms voltage step from −60 to +10 mV. B , retigabine (10 μM) significantly reduced inward Ca 2+ currents in isolated UBSM cells. C–E , isometric contractility of urinary bladder strips in response to 60 mM K + in the absence ( C ) or presence ( D ) of increasing concentrations of retigabine. Upward, square deflections in D are an artefact of tissue oxygenation and are not muscle contractions. Retigabine (1–10 μM) reduced steady‐state contractile responses to 60 mM K + as compared to vehicle (DMSO) ( E ). * P

    Journal: The Journal of Physiology

    Article Title: The KV7 channel activator retigabine suppresses mouse urinary bladder afferent nerve activity without affecting detrusor smooth muscle K+ channel currents

    doi: 10.1113/JP277021

    Figure Lengend Snippet: Retigabine inhibits Ca 2+ currents in freshly isolated mouse UBSM cells A , representative trace of inward Ca 2+ currents in response to a 20 ms voltage step from −60 to +10 mV. B , retigabine (10 μM) significantly reduced inward Ca 2+ currents in isolated UBSM cells. C–E , isometric contractility of urinary bladder strips in response to 60 mM K + in the absence ( C ) or presence ( D ) of increasing concentrations of retigabine. Upward, square deflections in D are an artefact of tissue oxygenation and are not muscle contractions. Retigabine (1–10 μM) reduced steady‐state contractile responses to 60 mM K + as compared to vehicle (DMSO) ( E ). * P

    Article Snippet: Retigabine was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Isolation, Mass Spectrometry

    Retigabine does not affect membrane potential in freshly isolated mouse UBSM cells A , representative trace of membrane potential of freshly isolated, current‐clamped UBSM cells in the presence of retigabine alone, retigabine with the K ATP channel opener pinacidil, and 140 mM extracellular K + . B , retigabine had no effect on membrane potential, whereas pinacidil caused hyperpolarization and 140 mM K + caused a reversible depolarization. Circles represent individual paired experiments. Bars represent mean ± SEM of each condition ( n = 3–7 cells from N = 3–5 mice).

    Journal: The Journal of Physiology

    Article Title: The KV7 channel activator retigabine suppresses mouse urinary bladder afferent nerve activity without affecting detrusor smooth muscle K+ channel currents

    doi: 10.1113/JP277021

    Figure Lengend Snippet: Retigabine does not affect membrane potential in freshly isolated mouse UBSM cells A , representative trace of membrane potential of freshly isolated, current‐clamped UBSM cells in the presence of retigabine alone, retigabine with the K ATP channel opener pinacidil, and 140 mM extracellular K + . B , retigabine had no effect on membrane potential, whereas pinacidil caused hyperpolarization and 140 mM K + caused a reversible depolarization. Circles represent individual paired experiments. Bars represent mean ± SEM of each condition ( n = 3–7 cells from N = 3–5 mice).

    Article Snippet: Retigabine was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Isolation, Mouse Assay

    Retigabine inhibits bursts of afferent activity during simulated TCs A–D , representative traces of intravesical pressure (top) and afferent nerve activity (bottom) during simulated TCs in the absence of drug ( A ) or in the presence of 10 μM retigabine ( B ), 10 μM XE‐991 ( C ), or both drugs ( D ). TCs were simulated by gentle compression of the bladder wall with plastic‐covered forceps. E , comparison of simulated TC peak afferent activity in the presence of retigabine, XE‐991, or a combination of both. Peak afferent activity per change in simulated TC integral was reduced by retigabine ( F ), an effect that was reversed by pre‐incubation of tissue with XE‐991. * P

    Journal: The Journal of Physiology

    Article Title: The KV7 channel activator retigabine suppresses mouse urinary bladder afferent nerve activity without affecting detrusor smooth muscle K+ channel currents

    doi: 10.1113/JP277021

    Figure Lengend Snippet: Retigabine inhibits bursts of afferent activity during simulated TCs A–D , representative traces of intravesical pressure (top) and afferent nerve activity (bottom) during simulated TCs in the absence of drug ( A ) or in the presence of 10 μM retigabine ( B ), 10 μM XE‐991 ( C ), or both drugs ( D ). TCs were simulated by gentle compression of the bladder wall with plastic‐covered forceps. E , comparison of simulated TC peak afferent activity in the presence of retigabine, XE‐991, or a combination of both. Peak afferent activity per change in simulated TC integral was reduced by retigabine ( F ), an effect that was reversed by pre‐incubation of tissue with XE‐991. * P

    Article Snippet: Retigabine was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Activity Assay, Incubation

    Retigabine inhibits all afferent nerve activity A–B , representative traces of intravesical pressure (top) and afferent nerve activity (bottom) in the presence of the L‐type Ca 2+ channel blocker diltiazem ( A ) or the K V 7 agonist retigabine ( B ). C , diltiazem (grey circles) blocked all transient contractions and associated bursts of nerve activity, but did not affect baseline afferent activity. Retigabine (black circles) not only abolished TCs and associated afferent activity, but also eliminated the remaining baseline afferent nerve activity. * P

    Journal: The Journal of Physiology

    Article Title: The KV7 channel activator retigabine suppresses mouse urinary bladder afferent nerve activity without affecting detrusor smooth muscle K+ channel currents

    doi: 10.1113/JP277021

    Figure Lengend Snippet: Retigabine inhibits all afferent nerve activity A–B , representative traces of intravesical pressure (top) and afferent nerve activity (bottom) in the presence of the L‐type Ca 2+ channel blocker diltiazem ( A ) or the K V 7 agonist retigabine ( B ). C , diltiazem (grey circles) blocked all transient contractions and associated bursts of nerve activity, but did not affect baseline afferent activity. Retigabine (black circles) not only abolished TCs and associated afferent activity, but also eliminated the remaining baseline afferent nerve activity. * P

    Article Snippet: Retigabine was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques: Activity Assay

    Retigabine‐sensitive inward currents are absent in the presence of 95 mM extracellular K + A , representative recordings of inward currents in response to the addition of 95 mM extracellular K + . B , summary graph showing that retigabine did not alter inward currents, whereas the K ATP channel opener pinacidil caused a significant inward K + current. Circles represent individual paired experiments. Bars represent mean ± SEM of each condition. * P

    Journal: The Journal of Physiology

    Article Title: The KV7 channel activator retigabine suppresses mouse urinary bladder afferent nerve activity without affecting detrusor smooth muscle K+ channel currents

    doi: 10.1113/JP277021

    Figure Lengend Snippet: Retigabine‐sensitive inward currents are absent in the presence of 95 mM extracellular K + A , representative recordings of inward currents in response to the addition of 95 mM extracellular K + . B , summary graph showing that retigabine did not alter inward currents, whereas the K ATP channel opener pinacidil caused a significant inward K + current. Circles represent individual paired experiments. Bars represent mean ± SEM of each condition. * P

    Article Snippet: Retigabine was obtained from Alomone Labs (Jerusalem, Israel).

    Techniques:

    HNa diet, in vivo: effects of an ETB-R antagonist on the ARNA responses to reflex increases in ERSNA.

    Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

    Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

    doi: 10.1152/ajpregu.91029.2008

    Figure Lengend Snippet: HNa diet, in vivo: effects of an ETB-R antagonist on the ARNA responses to reflex increases in ERSNA.

    Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

    Techniques: In Vivo

    Immunofluorescence double-labeling of T 13 DRG shows colocalization of ETB-R-like immunoreactivity (ETB-R-LI) with glial fibrillary acidic protein (GFAP)-LI (arrows) surrounding neuronal cell bodies ( a–c ). Also, many ETB-R-ir fiber-like structures

    Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

    Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

    doi: 10.1152/ajpregu.91029.2008

    Figure Lengend Snippet: Immunofluorescence double-labeling of T 13 DRG shows colocalization of ETB-R-like immunoreactivity (ETB-R-LI) with glial fibrillary acidic protein (GFAP)-LI (arrows) surrounding neuronal cell bodies ( a–c ). Also, many ETB-R-ir fiber-like structures

    Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

    Techniques: Immunofluorescence, Labeling

    Our studies suggest that ERSNA-induced increases in NE release lead to an increase in renal pelvic PGE 2 synthesis/release. In conditions of HNa dietary intake, activation of ETB-R on or close to unmyelinated Schwann cells surrounding the sensory nerves

    Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

    Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

    doi: 10.1152/ajpregu.91029.2008

    Figure Lengend Snippet: Our studies suggest that ERSNA-induced increases in NE release lead to an increase in renal pelvic PGE 2 synthesis/release. In conditions of HNa dietary intake, activation of ETB-R on or close to unmyelinated Schwann cells surrounding the sensory nerves

    Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

    Techniques: Activation Assay

    Immunofluorescence for ETB-R ( a ) in renal pelvic tissue shows nerve fiber-like structures (arrows) among smooth muscle cells. ETA-R-LI is present on smooth muscle cells in the renal pelvic wall and adjacent blood vessels (arrows, b ).

    Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

    Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

    doi: 10.1152/ajpregu.91029.2008

    Figure Lengend Snippet: Immunofluorescence for ETB-R ( a ) in renal pelvic tissue shows nerve fiber-like structures (arrows) among smooth muscle cells. ETA-R-LI is present on smooth muscle cells in the renal pelvic wall and adjacent blood vessels (arrows, b ).

    Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

    Techniques: Immunofluorescence

    Protein expression of ETB-R ( left ) and ETA-R ( right ) in T 9 –L 1 DRG and renal pelvic wall from rats fed the HNa and LNa diet. The ratio of ET-R/actin protein expression was normalized to the average value of the ratio in HNa diet rats. *

    Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

    Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

    doi: 10.1152/ajpregu.91029.2008

    Figure Lengend Snippet: Protein expression of ETB-R ( left ) and ETA-R ( right ) in T 9 –L 1 DRG and renal pelvic wall from rats fed the HNa and LNa diet. The ratio of ET-R/actin protein expression was normalized to the average value of the ratio in HNa diet rats. *

    Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

    Techniques: Expressing

    A : in vivo, effects of renal pelvic administration of the ETB-R antagonist BQ788 on the ERSNA and ARNA responses to placing the rat's tail in 47°C water in rats fed HNa diet. B : effects of renal pelvic administration of the ETA-R antagonist BQ123

    Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

    Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

    doi: 10.1152/ajpregu.91029.2008

    Figure Lengend Snippet: A : in vivo, effects of renal pelvic administration of the ETB-R antagonist BQ788 on the ERSNA and ARNA responses to placing the rat's tail in 47°C water in rats fed HNa diet. B : effects of renal pelvic administration of the ETA-R antagonist BQ123

    Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

    Techniques: In Vivo

    Immunofluorescence double-labeling of renal pelvic tissue shows ETB-R-ir fiber-like structures among smooth muscle cells ( a–c ) close to calcitonin gene-related peptide-immunoreactive (CGRP-ir) sensory nerve fibers ( d–f ). αSMA,

    Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

    Article Title: Dietary sodium modulates the interaction between efferent renal sympathetic nerve activity and afferent renal nerve activity: role of endothelin

    doi: 10.1152/ajpregu.91029.2008

    Figure Lengend Snippet: Immunofluorescence double-labeling of renal pelvic tissue shows ETB-R-ir fiber-like structures among smooth muscle cells ( a–c ) close to calcitonin gene-related peptide-immunoreactive (CGRP-ir) sensory nerve fibers ( d–f ). αSMA,

    Article Snippet: After completion of the protocol for TSA for detection of GFAP or CGRP, the tissue sections were incubated with antiserum for ETB-R (rabbit; 1:100, Alomone Labs) and processed by the indirect immunofluorescence technique.

    Techniques: Immunofluorescence, Labeling

    NT-3 generated long-range Ca 2+ signaling from the axon to the cell body. a Long-range Ca 2+ wave. The relative change in the Cal-520 emission ratio (defined as R) was used as a measure of changes in Ca 2+ concentration. The pseudocolored images represent R after local application of PBS ( top ) or NT-3 ( bottom ) to the axon ( arrow ). Scale bars, 50 μm. b The mean amplitude of R treat / R 0 for 90 s during local application of NT-3 in the presence of the indicated inhibitors (PBS = 10, NT-3 = 16, xestospongin C = 9, ryanodine = 15, dantrolene = 18, SKF96365 = 12 neurons from three independent experiments). c , d The axon was exposed to NT-3 in the presence of the indicated inhibitors, and then minor neurite outgrowth (PBS = 21, NT-3 = 21, xestospongin C = 26, ryanodine = 25, dantrolene = 25, SKF96365 = 24 neurites from three independent experiments) c and axonal outgrowth (PBS = 7, NT-3 = 9, xestospongin C = 9, ryanodine = 9, dantrolene = 8, SKF96365 = 8 neurons from three independent experiments) d were measured. e Local application of NT-3 increased the quantity of phospho-CaMKI in the cell body. After local application of PBS ( top ) or NT-3 ( bottom ), hippocampal neurons were immunostained with antibodies against CaMKI ( green ) and phospho-Thr177 of CaMKI ( magenta ). The merged images ( right panels ) are shown. The graph plots the fluorescence intensities of total CaMKI ( green ) and CaMKI phosphorylated at Thr177 ( magenta ) and in the line. Scale bars, 20 μm. f NT-3-induced minor neurite retraction was abolished by Ca 2+ signaling inhibitors. The axon was exposed to NT-3 in the presence of the indicated inhibitors, and minor neurite outgrowth was measured (PBS = 27, NT-3 = 31, BAPTA = 47, STO-609 = 45, KN-93 = 40 neurites from three independent experiments). g , h Local application of indicated inhibitors to the axon. Minor neurite outgrowth (DMSO = 42, xestospongin C = 32, ryanodine = 37, dantrolene = 35, SKF96365 = 32 neurons from three independent experiments) g and axonal outgrowth (DMSO = 14, xestospongin C = 11, ryanodine = 13, dantrolene = 13, SKF96365 = 12 neurons from three independent experiments) h were measured. Error bars represent SEM. * P

    Journal: Nature Communications

    Article Title: Discovery of long-range inhibitory signaling to ensure single axon formation

    doi: 10.1038/s41467-017-00044-2

    Figure Lengend Snippet: NT-3 generated long-range Ca 2+ signaling from the axon to the cell body. a Long-range Ca 2+ wave. The relative change in the Cal-520 emission ratio (defined as R) was used as a measure of changes in Ca 2+ concentration. The pseudocolored images represent R after local application of PBS ( top ) or NT-3 ( bottom ) to the axon ( arrow ). Scale bars, 50 μm. b The mean amplitude of R treat / R 0 for 90 s during local application of NT-3 in the presence of the indicated inhibitors (PBS = 10, NT-3 = 16, xestospongin C = 9, ryanodine = 15, dantrolene = 18, SKF96365 = 12 neurons from three independent experiments). c , d The axon was exposed to NT-3 in the presence of the indicated inhibitors, and then minor neurite outgrowth (PBS = 21, NT-3 = 21, xestospongin C = 26, ryanodine = 25, dantrolene = 25, SKF96365 = 24 neurites from three independent experiments) c and axonal outgrowth (PBS = 7, NT-3 = 9, xestospongin C = 9, ryanodine = 9, dantrolene = 8, SKF96365 = 8 neurons from three independent experiments) d were measured. e Local application of NT-3 increased the quantity of phospho-CaMKI in the cell body. After local application of PBS ( top ) or NT-3 ( bottom ), hippocampal neurons were immunostained with antibodies against CaMKI ( green ) and phospho-Thr177 of CaMKI ( magenta ). The merged images ( right panels ) are shown. The graph plots the fluorescence intensities of total CaMKI ( green ) and CaMKI phosphorylated at Thr177 ( magenta ) and in the line. Scale bars, 20 μm. f NT-3-induced minor neurite retraction was abolished by Ca 2+ signaling inhibitors. The axon was exposed to NT-3 in the presence of the indicated inhibitors, and minor neurite outgrowth was measured (PBS = 27, NT-3 = 31, BAPTA = 47, STO-609 = 45, KN-93 = 40 neurites from three independent experiments). g , h Local application of indicated inhibitors to the axon. Minor neurite outgrowth (DMSO = 42, xestospongin C = 32, ryanodine = 37, dantrolene = 35, SKF96365 = 32 neurons from three independent experiments) g and axonal outgrowth (DMSO = 14, xestospongin C = 11, ryanodine = 13, dantrolene = 13, SKF96365 = 12 neurons from three independent experiments) h were measured. Error bars represent SEM. * P

    Article Snippet: In some experiments, the following reagents were applied to the culture medium at least 30 min before the application of the NT-3 gradients as previouslydescribed : 2 μM xestospongin C, 20 μM dantrolene, 3 μM SKF96365, 100 μM ryanodine (Alomone Labs), 5 μM BAPTA-AM (Invitrogen), 1 μM KN-93 or 5 μM STO-609.

    Techniques: Generated, Concentration Assay, Fluorescence