ω conotoxin gvia  (Alomone Labs)


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    Alomone Labs ω conotoxin gvia
    ( A ) Isolated magnocellular neurons were identified as either vasopressin or oxytocin by their presence or absence of the GFP signal. Scale bar = 10 µm. ( B ) The current-voltage relationships show no difference between whole-cell calcium currents (WCCC) of vasopressin and oxytocin neurons (n = 20 each). ( C ) Normalized conductance (G/G max , fitted using Boltzmann equation) showed no significant change with or without TG. ( D ) Examples of two neurons (both oxytocin neurons), one vehicle treated (•) and one 50 min after pre-treating with thapsigargin (○). The steady-state Ca 2+ current is plotted against time following treatment with channel toxins (in order: 1 µM nicardipine; 200 nM ω-agatoxin IV; 500 <t>nM</t> <t>ω-conotoxin</t> <t>GVIA).</t> The current measured after each toxin was subtracted from that evoked before each treatment to determine the current carried by L-, P/Q- and N-type channels. The remaining current was attributed to R-type channels. ( E ) Examples of N-, L- and WCCC elicited by voltage steps in controls and TG treated cells. ( F ) The resulting current density for each current type normalised for the peak maximal current was averaged (n = 5) to give the average current density from neurons treated with vehicle or thapsigargin. ( G ) In a subsequent experiment L- or N-type channel toxins were administered, alternating the order (n = 5 for each condition). Mean±S.E.M are shown and compared by t-test. * P <0.05 vs control.
    ω Conotoxin Gvia, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "The Involvement of Voltage-Operated Calcium Channels in Somato-Dendritic Oxytocin Release"

    Article Title: The Involvement of Voltage-Operated Calcium Channels in Somato-Dendritic Oxytocin Release

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0025366

    ( A ) Isolated magnocellular neurons were identified as either vasopressin or oxytocin by their presence or absence of the GFP signal. Scale bar = 10 µm. ( B ) The current-voltage relationships show no difference between whole-cell calcium currents (WCCC) of vasopressin and oxytocin neurons (n = 20 each). ( C ) Normalized conductance (G/G max , fitted using Boltzmann equation) showed no significant change with or without TG. ( D ) Examples of two neurons (both oxytocin neurons), one vehicle treated (•) and one 50 min after pre-treating with thapsigargin (○). The steady-state Ca 2+ current is plotted against time following treatment with channel toxins (in order: 1 µM nicardipine; 200 nM ω-agatoxin IV; 500 nM ω-conotoxin GVIA). The current measured after each toxin was subtracted from that evoked before each treatment to determine the current carried by L-, P/Q- and N-type channels. The remaining current was attributed to R-type channels. ( E ) Examples of N-, L- and WCCC elicited by voltage steps in controls and TG treated cells. ( F ) The resulting current density for each current type normalised for the peak maximal current was averaged (n = 5) to give the average current density from neurons treated with vehicle or thapsigargin. ( G ) In a subsequent experiment L- or N-type channel toxins were administered, alternating the order (n = 5 for each condition). Mean±S.E.M are shown and compared by t-test. * P <0.05 vs control.
    Figure Legend Snippet: ( A ) Isolated magnocellular neurons were identified as either vasopressin or oxytocin by their presence or absence of the GFP signal. Scale bar = 10 µm. ( B ) The current-voltage relationships show no difference between whole-cell calcium currents (WCCC) of vasopressin and oxytocin neurons (n = 20 each). ( C ) Normalized conductance (G/G max , fitted using Boltzmann equation) showed no significant change with or without TG. ( D ) Examples of two neurons (both oxytocin neurons), one vehicle treated (•) and one 50 min after pre-treating with thapsigargin (○). The steady-state Ca 2+ current is plotted against time following treatment with channel toxins (in order: 1 µM nicardipine; 200 nM ω-agatoxin IV; 500 nM ω-conotoxin GVIA). The current measured after each toxin was subtracted from that evoked before each treatment to determine the current carried by L-, P/Q- and N-type channels. The remaining current was attributed to R-type channels. ( E ) Examples of N-, L- and WCCC elicited by voltage steps in controls and TG treated cells. ( F ) The resulting current density for each current type normalised for the peak maximal current was averaged (n = 5) to give the average current density from neurons treated with vehicle or thapsigargin. ( G ) In a subsequent experiment L- or N-type channel toxins were administered, alternating the order (n = 5 for each condition). Mean±S.E.M are shown and compared by t-test. * P <0.05 vs control.

    Techniques Used: Isolation

    ω conotoxin gvia  (Alomone Labs)


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    Alomone Labs ω conotoxin gvia
    ( A ) Isolated magnocellular neurons were identified as either vasopressin or oxytocin by their presence or absence of the GFP signal. Scale bar = 10 µm. ( B ) The current-voltage relationships show no difference between whole-cell calcium currents (WCCC) of vasopressin and oxytocin neurons (n = 20 each). ( C ) Normalized conductance (G/G max , fitted using Boltzmann equation) showed no significant change with or without TG. ( D ) Examples of two neurons (both oxytocin neurons), one vehicle treated (•) and one 50 min after pre-treating with thapsigargin (○). The steady-state Ca 2+ current is plotted against time following treatment with channel toxins (in order: 1 µM nicardipine; 200 nM ω-agatoxin IV; 500 <t>nM</t> <t>ω-conotoxin</t> <t>GVIA).</t> The current measured after each toxin was subtracted from that evoked before each treatment to determine the current carried by L-, P/Q- and N-type channels. The remaining current was attributed to R-type channels. ( E ) Examples of N-, L- and WCCC elicited by voltage steps in controls and TG treated cells. ( F ) The resulting current density for each current type normalised for the peak maximal current was averaged (n = 5) to give the average current density from neurons treated with vehicle or thapsigargin. ( G ) In a subsequent experiment L- or N-type channel toxins were administered, alternating the order (n = 5 for each condition). Mean±S.E.M are shown and compared by t-test. * P <0.05 vs control.
    ω Conotoxin Gvia, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "The Involvement of Voltage-Operated Calcium Channels in Somato-Dendritic Oxytocin Release"

    Article Title: The Involvement of Voltage-Operated Calcium Channels in Somato-Dendritic Oxytocin Release

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0025366

    ( A ) Isolated magnocellular neurons were identified as either vasopressin or oxytocin by their presence or absence of the GFP signal. Scale bar = 10 µm. ( B ) The current-voltage relationships show no difference between whole-cell calcium currents (WCCC) of vasopressin and oxytocin neurons (n = 20 each). ( C ) Normalized conductance (G/G max , fitted using Boltzmann equation) showed no significant change with or without TG. ( D ) Examples of two neurons (both oxytocin neurons), one vehicle treated (•) and one 50 min after pre-treating with thapsigargin (○). The steady-state Ca 2+ current is plotted against time following treatment with channel toxins (in order: 1 µM nicardipine; 200 nM ω-agatoxin IV; 500 nM ω-conotoxin GVIA). The current measured after each toxin was subtracted from that evoked before each treatment to determine the current carried by L-, P/Q- and N-type channels. The remaining current was attributed to R-type channels. ( E ) Examples of N-, L- and WCCC elicited by voltage steps in controls and TG treated cells. ( F ) The resulting current density for each current type normalised for the peak maximal current was averaged (n = 5) to give the average current density from neurons treated with vehicle or thapsigargin. ( G ) In a subsequent experiment L- or N-type channel toxins were administered, alternating the order (n = 5 for each condition). Mean±S.E.M are shown and compared by t-test. * P <0.05 vs control.
    Figure Legend Snippet: ( A ) Isolated magnocellular neurons were identified as either vasopressin or oxytocin by their presence or absence of the GFP signal. Scale bar = 10 µm. ( B ) The current-voltage relationships show no difference between whole-cell calcium currents (WCCC) of vasopressin and oxytocin neurons (n = 20 each). ( C ) Normalized conductance (G/G max , fitted using Boltzmann equation) showed no significant change with or without TG. ( D ) Examples of two neurons (both oxytocin neurons), one vehicle treated (•) and one 50 min after pre-treating with thapsigargin (○). The steady-state Ca 2+ current is plotted against time following treatment with channel toxins (in order: 1 µM nicardipine; 200 nM ω-agatoxin IV; 500 nM ω-conotoxin GVIA). The current measured after each toxin was subtracted from that evoked before each treatment to determine the current carried by L-, P/Q- and N-type channels. The remaining current was attributed to R-type channels. ( E ) Examples of N-, L- and WCCC elicited by voltage steps in controls and TG treated cells. ( F ) The resulting current density for each current type normalised for the peak maximal current was averaged (n = 5) to give the average current density from neurons treated with vehicle or thapsigargin. ( G ) In a subsequent experiment L- or N-type channel toxins were administered, alternating the order (n = 5 for each condition). Mean±S.E.M are shown and compared by t-test. * P <0.05 vs control.

    Techniques Used: Isolation

    conotoxin gvia  (Alomone Labs)


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    Alomone Labs conotoxin gvia
    Summary of the compounds and antiepileptic drugs evaluated in the calcium oscillations assay.
    Conotoxin Gvia, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "New In Vitro Phenotypic Assay for Epilepsy: Fluorescent Measurement of Synchronized Neuronal Calcium Oscillations"

    Article Title: New In Vitro Phenotypic Assay for Epilepsy: Fluorescent Measurement of Synchronized Neuronal Calcium Oscillations

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0084755

    Summary of the compounds and antiepileptic drugs evaluated in the calcium oscillations assay.
    Figure Legend Snippet: Summary of the compounds and antiepileptic drugs evaluated in the calcium oscillations assay.

    Techniques Used:

    ω conotoxin gvia ω gvia  (Alomone Labs)


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    Alomone Labs ω conotoxin gvia ω gvia
    SPC-01 generates functional neurons. (a) Preincubation with Cd 2+ (100 μ M ) together with Ni 2+ (50 μ M ), for 5 minutes significantly reduced the [Ca 2+ ] i responses induced by K + in all cells tested by 93% ± 9.2% ( n = 5), indicating the involvement of voltage-activated Ca 2+ channels in depolarization-induced Ca 2+ entry. (b) The specific L-type Ca 2+ channel blocker nicardipine (1 μ M ) reduced the [Ca 2+ ] i responses by 28% ± 7% ( n = 5; P = 0.002). The trace in (c) shows the [Ca 2+ ] i responses observed in a selected SPC-01-derived neuron induced by 50 m M K + , preincubated for 2 minutes with <t>Ω</t> <t>-GVIA</t> (800 n M ) and then stimulated with K + . After a washout of 10 minutes, the same cells were subjected to K + to observe recovery. Similarly, the effect of the P/Q-Ca 2+ channel blocker, Ω -Aga-IVA, 300 n M was tested before and during stimulation with high K + (d) . After washout of the toxin, the [Ca 2+ ] i response was shown to be reversible. The histogram (e) summarizes the results presented in (c) and (d) . The resting [Ca 2+ ] i level in these cells is indicated as basal. The results are expressed as mean ± SEM; n = 4 (Ω-GVIA) and n = 5 (Ω-Aga-IVA). Asterisks indicate the statistical significance ( P > 0.05) versus control K + stimulus.
    ω Conotoxin Gvia ω Gvia, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Conditionally immortalized stem cell lines from human spinal cord retain regional identity and generate functional V2a interneurons and motorneurons"

    Article Title: Conditionally immortalized stem cell lines from human spinal cord retain regional identity and generate functional V2a interneurons and motorneurons

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/scrt220

    SPC-01 generates functional neurons. (a) Preincubation with Cd 2+ (100 μ M ) together with Ni 2+ (50 μ M ), for 5 minutes significantly reduced the [Ca 2+ ] i responses induced by K + in all cells tested by 93% ± 9.2% ( n = 5), indicating the involvement of voltage-activated Ca 2+ channels in depolarization-induced Ca 2+ entry. (b) The specific L-type Ca 2+ channel blocker nicardipine (1 μ M ) reduced the [Ca 2+ ] i responses by 28% ± 7% ( n = 5; P = 0.002). The trace in (c) shows the [Ca 2+ ] i responses observed in a selected SPC-01-derived neuron induced by 50 m M K + , preincubated for 2 minutes with Ω -GVIA (800 n M ) and then stimulated with K + . After a washout of 10 minutes, the same cells were subjected to K + to observe recovery. Similarly, the effect of the P/Q-Ca 2+ channel blocker, Ω -Aga-IVA, 300 n M was tested before and during stimulation with high K + (d) . After washout of the toxin, the [Ca 2+ ] i response was shown to be reversible. The histogram (e) summarizes the results presented in (c) and (d) . The resting [Ca 2+ ] i level in these cells is indicated as basal. The results are expressed as mean ± SEM; n = 4 (Ω-GVIA) and n = 5 (Ω-Aga-IVA). Asterisks indicate the statistical significance ( P > 0.05) versus control K + stimulus.
    Figure Legend Snippet: SPC-01 generates functional neurons. (a) Preincubation with Cd 2+ (100 μ M ) together with Ni 2+ (50 μ M ), for 5 minutes significantly reduced the [Ca 2+ ] i responses induced by K + in all cells tested by 93% ± 9.2% ( n = 5), indicating the involvement of voltage-activated Ca 2+ channels in depolarization-induced Ca 2+ entry. (b) The specific L-type Ca 2+ channel blocker nicardipine (1 μ M ) reduced the [Ca 2+ ] i responses by 28% ± 7% ( n = 5; P = 0.002). The trace in (c) shows the [Ca 2+ ] i responses observed in a selected SPC-01-derived neuron induced by 50 m M K + , preincubated for 2 minutes with Ω -GVIA (800 n M ) and then stimulated with K + . After a washout of 10 minutes, the same cells were subjected to K + to observe recovery. Similarly, the effect of the P/Q-Ca 2+ channel blocker, Ω -Aga-IVA, 300 n M was tested before and during stimulation with high K + (d) . After washout of the toxin, the [Ca 2+ ] i response was shown to be reversible. The histogram (e) summarizes the results presented in (c) and (d) . The resting [Ca 2+ ] i level in these cells is indicated as basal. The results are expressed as mean ± SEM; n = 4 (Ω-GVIA) and n = 5 (Ω-Aga-IVA). Asterisks indicate the statistical significance ( P > 0.05) versus control K + stimulus.

    Techniques Used: Functional Assay, Derivative Assay

    ω conotoxin gvia  (Alomone Labs)


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    Alomone Labs ω conotoxin gvia
    ω Conotoxin Gvia, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ω conotoxin gvia  (Alomone Labs)


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    Alomone Labs ω conotoxin gvia
    ω Conotoxin Gvia, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ω conotoxin gvia  (Alomone Labs)


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    Alomone Labs ω conotoxin gvia
    A , Left , Time courses of I Ca amplitude during serial application of nifedipine (10 µM), ω-agatoxin IVA (0.5 <t>µM),</t> <t>ω-conotoxin</t> <t>GVIA</t> (3 µM), SNX-482 (300 nM) and CdCl 2 (100 µM). I Ca was evoked every 10 s by 70 ms test pulses to 0 mV from a holding potential of −80 mV. The horizontal bars indicate the duration of drug application. Right , superimposed current traces obtained at different time points during drug application (labeled as a–f). B , Bar graph representing the mean I Ca inhibition (%) produced by application of the indicated antagonists or toxins. Error bars represent s . e . m . The number of neurons tested is indicated in parentheses. C , Effect of non-dihydropyridine Ca 2+ channel agonist FPL 64176 (FPL) on I Ca in DRG neurons. FPL (1 µM) was applied to rat superior cervical ganglion (SCG) neurons as a positive control (left panel). Note that FPL applied to zebrafish DRG I Ca display neither an increase in macroscopic inward currents nor greatly prolonged trajectory of the tail currents (right panel).
    ω Conotoxin Gvia, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Characterization of Na + and Ca 2+ Channels in Zebrafish Dorsal Root Ganglion Neurons"

    Article Title: Characterization of Na + and Ca 2+ Channels in Zebrafish Dorsal Root Ganglion Neurons

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0042602

    A , Left , Time courses of I Ca amplitude during serial application of nifedipine (10 µM), ω-agatoxin IVA (0.5 µM), ω-conotoxin GVIA (3 µM), SNX-482 (300 nM) and CdCl 2 (100 µM). I Ca was evoked every 10 s by 70 ms test pulses to 0 mV from a holding potential of −80 mV. The horizontal bars indicate the duration of drug application. Right , superimposed current traces obtained at different time points during drug application (labeled as a–f). B , Bar graph representing the mean I Ca inhibition (%) produced by application of the indicated antagonists or toxins. Error bars represent s . e . m . The number of neurons tested is indicated in parentheses. C , Effect of non-dihydropyridine Ca 2+ channel agonist FPL 64176 (FPL) on I Ca in DRG neurons. FPL (1 µM) was applied to rat superior cervical ganglion (SCG) neurons as a positive control (left panel). Note that FPL applied to zebrafish DRG I Ca display neither an increase in macroscopic inward currents nor greatly prolonged trajectory of the tail currents (right panel).
    Figure Legend Snippet: A , Left , Time courses of I Ca amplitude during serial application of nifedipine (10 µM), ω-agatoxin IVA (0.5 µM), ω-conotoxin GVIA (3 µM), SNX-482 (300 nM) and CdCl 2 (100 µM). I Ca was evoked every 10 s by 70 ms test pulses to 0 mV from a holding potential of −80 mV. The horizontal bars indicate the duration of drug application. Right , superimposed current traces obtained at different time points during drug application (labeled as a–f). B , Bar graph representing the mean I Ca inhibition (%) produced by application of the indicated antagonists or toxins. Error bars represent s . e . m . The number of neurons tested is indicated in parentheses. C , Effect of non-dihydropyridine Ca 2+ channel agonist FPL 64176 (FPL) on I Ca in DRG neurons. FPL (1 µM) was applied to rat superior cervical ganglion (SCG) neurons as a positive control (left panel). Note that FPL applied to zebrafish DRG I Ca display neither an increase in macroscopic inward currents nor greatly prolonged trajectory of the tail currents (right panel).

    Techniques Used: Labeling, Inhibition, Produced, Positive Control

    ω conotoxin gvia  (Alomone Labs)


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    Alomone Labs ω conotoxin gvia
    Pharmacological and biophysical dissection of TAT-CBD3A6K-mediated block of T-and R-type calcium currents in DRG neurons. ( A ) Representative T- (top) and R-type (bottom) current traces obtained from two separate DRG neurons evoked by 200 ms steps in 5 mV increments from −60 mV to +50 mV, from a holding potential of −90 mV. The extracellular bath solution contained 5 mM Nifedipine (Nif), 200 nM ω-Agatoxin IVA (Aga) and 500 <t>nM</t> <t>ω-Conotoxin</t> <t>GVIA</t> (CTX) to block L-, P/Q-, and N-type calcium currents, respectively. ( B ) Summary of the normalized conductance (G) versus voltage relations for DRG neurons with T- (filled squares) or R- (open squares) type calcium currents. The dotted line at −10 mV highlights the clear discrimination in conductances between T- and R-type currents. ( C ) Representative currents, evoked by a ramp depolarizations from −60 mV to +20 mV for 2 s, illustrating the presence of both T- and R-type currents in the same DRG neuron before (left trace) and 2 min after application 10 μM TAT-CBD3A6K (right trace)
    ω Conotoxin Gvia, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "CRMP-2 peptide mediated decrease of high and low voltage-activated calcium channels, attenuation of nociceptor excitability, and anti-nociception in a model of AIDS therapy-induced painful peripheral neuropathy"

    Article Title: CRMP-2 peptide mediated decrease of high and low voltage-activated calcium channels, attenuation of nociceptor excitability, and anti-nociception in a model of AIDS therapy-induced painful peripheral neuropathy

    Journal: Molecular Pain

    doi: 10.1186/1744-8069-8-54

    Pharmacological and biophysical dissection of TAT-CBD3A6K-mediated block of T-and R-type calcium currents in DRG neurons. ( A ) Representative T- (top) and R-type (bottom) current traces obtained from two separate DRG neurons evoked by 200 ms steps in 5 mV increments from −60 mV to +50 mV, from a holding potential of −90 mV. The extracellular bath solution contained 5 mM Nifedipine (Nif), 200 nM ω-Agatoxin IVA (Aga) and 500 nM ω-Conotoxin GVIA (CTX) to block L-, P/Q-, and N-type calcium currents, respectively. ( B ) Summary of the normalized conductance (G) versus voltage relations for DRG neurons with T- (filled squares) or R- (open squares) type calcium currents. The dotted line at −10 mV highlights the clear discrimination in conductances between T- and R-type currents. ( C ) Representative currents, evoked by a ramp depolarizations from −60 mV to +20 mV for 2 s, illustrating the presence of both T- and R-type currents in the same DRG neuron before (left trace) and 2 min after application 10 μM TAT-CBD3A6K (right trace)
    Figure Legend Snippet: Pharmacological and biophysical dissection of TAT-CBD3A6K-mediated block of T-and R-type calcium currents in DRG neurons. ( A ) Representative T- (top) and R-type (bottom) current traces obtained from two separate DRG neurons evoked by 200 ms steps in 5 mV increments from −60 mV to +50 mV, from a holding potential of −90 mV. The extracellular bath solution contained 5 mM Nifedipine (Nif), 200 nM ω-Agatoxin IVA (Aga) and 500 nM ω-Conotoxin GVIA (CTX) to block L-, P/Q-, and N-type calcium currents, respectively. ( B ) Summary of the normalized conductance (G) versus voltage relations for DRG neurons with T- (filled squares) or R- (open squares) type calcium currents. The dotted line at −10 mV highlights the clear discrimination in conductances between T- and R-type currents. ( C ) Representative currents, evoked by a ramp depolarizations from −60 mV to +20 mV for 2 s, illustrating the presence of both T- and R-type currents in the same DRG neuron before (left trace) and 2 min after application 10 μM TAT-CBD3A6K (right trace)

    Techniques Used: Dissection, Blocking Assay

    Characterization of TAT-CBD3A6K-mediated inhibition of T- and R-type calcium currents. ( A ) Representative family of traces from a DRG neuron with both T- and R-type calcium currents before ( left ), 2 min ( middle ) and 5 min ( right ) after addition of 10 μM TAT-CBD3A6K. Currents were elicited in response to the voltage protocol described in the legend to Figure A. To isolate T- and R-type calcium currents, the extracellular bath solution contained 5 mM Nifedipine (Nif), 200 nM ω-Agatoxin IVA (Aga) and 500 nM ω-Conotoxin GVIA (CTX) to block L-, P/Q-, and N-type calcium currents, respectively. ( B , C ) Time course of TAT-CBD3A6K mediated inhibition (“run-down”) of T-type ( B ) and R-type ( C ) calcium currents. Time course of inhibition is shown as averaged normalized current density (pA pF -1 ) before peptide addition and at intervals of 30 s for 5 min. Averaged values are shown with standard error for 4–6 control cells and 4 cells following addition of 10 μM TAT-CBD3A6K. The asterisk denotes statistical significance (p < 0.05; Student’s t -test) compared to the corresponding control time point. Some error bars are smaller than the symbols. Data represent mean ± SEM from n = 3–6 cells at each time point except for n = 2 the 4 min time point for T-type currents in the presence of peptide
    Figure Legend Snippet: Characterization of TAT-CBD3A6K-mediated inhibition of T- and R-type calcium currents. ( A ) Representative family of traces from a DRG neuron with both T- and R-type calcium currents before ( left ), 2 min ( middle ) and 5 min ( right ) after addition of 10 μM TAT-CBD3A6K. Currents were elicited in response to the voltage protocol described in the legend to Figure A. To isolate T- and R-type calcium currents, the extracellular bath solution contained 5 mM Nifedipine (Nif), 200 nM ω-Agatoxin IVA (Aga) and 500 nM ω-Conotoxin GVIA (CTX) to block L-, P/Q-, and N-type calcium currents, respectively. ( B , C ) Time course of TAT-CBD3A6K mediated inhibition (“run-down”) of T-type ( B ) and R-type ( C ) calcium currents. Time course of inhibition is shown as averaged normalized current density (pA pF -1 ) before peptide addition and at intervals of 30 s for 5 min. Averaged values are shown with standard error for 4–6 control cells and 4 cells following addition of 10 μM TAT-CBD3A6K. The asterisk denotes statistical significance (p < 0.05; Student’s t -test) compared to the corresponding control time point. Some error bars are smaller than the symbols. Data represent mean ± SEM from n = 3–6 cells at each time point except for n = 2 the 4 min time point for T-type currents in the presence of peptide

    Techniques Used: Inhibition, Blocking Assay

    ω conotoxin gvia  (Alomone Labs)


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    Alomone Labs ω conotoxin gvia
    Effect of VGCC blockers on the Ca 2+ response of the cells to a 150 ns pulse. Results are plotted as the averaged cell responses ± SE for cells exposed to a 3.1 kV/cm pulse in the absence and presence of (a) 100 nM ω -agatoxin IVA, ( n = 6), (b) 20 nM <t>ω</t> <t>-conotoxin</t> <t>GVIA,</t> ( n = 8–14) (c), 5 μ M nitrendipine, ( n = 9–12), and (d) a cocktail of all three blockers, ( n = 11–13). The arrow indicates when the pulse was delivered to the cells. Cell were pretreated with the blockers for 1 hr at room temperature. ∗ p < 0.05, significantly different from control.
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    1) Product Images from "Different Membrane Pathways Mediate Ca 2+ Influx in Adrenal Chromaffin Cells Exposed to 150–400 ns Electric Pulses"

    Article Title: Different Membrane Pathways Mediate Ca 2+ Influx in Adrenal Chromaffin Cells Exposed to 150–400 ns Electric Pulses

    Journal: BioMed Research International

    doi: 10.1155/2018/9046891

    Effect of VGCC blockers on the Ca 2+ response of the cells to a 150 ns pulse. Results are plotted as the averaged cell responses ± SE for cells exposed to a 3.1 kV/cm pulse in the absence and presence of (a) 100 nM ω -agatoxin IVA, ( n = 6), (b) 20 nM ω -conotoxin GVIA, ( n = 8–14) (c), 5 μ M nitrendipine, ( n = 9–12), and (d) a cocktail of all three blockers, ( n = 11–13). The arrow indicates when the pulse was delivered to the cells. Cell were pretreated with the blockers for 1 hr at room temperature. ∗ p < 0.05, significantly different from control.
    Figure Legend Snippet: Effect of VGCC blockers on the Ca 2+ response of the cells to a 150 ns pulse. Results are plotted as the averaged cell responses ± SE for cells exposed to a 3.1 kV/cm pulse in the absence and presence of (a) 100 nM ω -agatoxin IVA, ( n = 6), (b) 20 nM ω -conotoxin GVIA, ( n = 8–14) (c), 5 μ M nitrendipine, ( n = 9–12), and (d) a cocktail of all three blockers, ( n = 11–13). The arrow indicates when the pulse was delivered to the cells. Cell were pretreated with the blockers for 1 hr at room temperature. ∗ p < 0.05, significantly different from control.

    Techniques Used:

    Effect of blocking VGCCs on 150 ns pulse-induced increases in [Ca 2+ ] i .
    Figure Legend Snippet: Effect of blocking VGCCs on 150 ns pulse-induced increases in [Ca 2+ ] i .

    Techniques Used: Blocking Assay, Fluorescence

    ω conotoxin gvia c 300  (Alomone Labs)


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    Alomone Labs ω conotoxin gvia c 300
    The effect of eupatilin in combination <t>with</t> <t>ω-conotoxin</t> <t>GVIA</t> or ω-agatoxin IVA on 4-AP-induced release of glutamate in rat cerebral cortex synaptosomes. Eupatilin was added 10 min before 4-AP addition and ω-conotoxin GVIA or ω-agatoxin IVA was added 20 min before this. Data are the mean ± SEM. ***, p < 0.001, as compared to the control; #, p < 0.05, as compared to the ω-conotoxin GVIA-treated group; n = 5 per group.
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    1) Product Images from "Inhibition of Synaptic Glutamate Exocytosis and Prevention of Glutamate Neurotoxicity by Eupatilin from Artemisia argyi in the Rat Cortex"

    Article Title: Inhibition of Synaptic Glutamate Exocytosis and Prevention of Glutamate Neurotoxicity by Eupatilin from Artemisia argyi in the Rat Cortex

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms232113406

    The effect of eupatilin in combination with ω-conotoxin GVIA or ω-agatoxin IVA on 4-AP-induced release of glutamate in rat cerebral cortex synaptosomes. Eupatilin was added 10 min before 4-AP addition and ω-conotoxin GVIA or ω-agatoxin IVA was added 20 min before this. Data are the mean ± SEM. ***, p < 0.001, as compared to the control; #, p < 0.05, as compared to the ω-conotoxin GVIA-treated group; n = 5 per group.
    Figure Legend Snippet: The effect of eupatilin in combination with ω-conotoxin GVIA or ω-agatoxin IVA on 4-AP-induced release of glutamate in rat cerebral cortex synaptosomes. Eupatilin was added 10 min before 4-AP addition and ω-conotoxin GVIA or ω-agatoxin IVA was added 20 min before this. Data are the mean ± SEM. ***, p < 0.001, as compared to the control; #, p < 0.05, as compared to the ω-conotoxin GVIA-treated group; n = 5 per group.

    Techniques Used:

    ω conotoxin gvia  (Alomone Labs)


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    ω Conotoxin Gvia, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs ω conotoxin gvia
    ( A ) Isolated magnocellular neurons were identified as either vasopressin or oxytocin by their presence or absence of the GFP signal. Scale bar = 10 µm. ( B ) The current-voltage relationships show no difference between whole-cell calcium currents (WCCC) of vasopressin and oxytocin neurons (n = 20 each). ( C ) Normalized conductance (G/G max , fitted using Boltzmann equation) showed no significant change with or without TG. ( D ) Examples of two neurons (both oxytocin neurons), one vehicle treated (•) and one 50 min after pre-treating with thapsigargin (○). The steady-state Ca 2+ current is plotted against time following treatment with channel toxins (in order: 1 µM nicardipine; 200 nM ω-agatoxin IV; 500 <t>nM</t> <t>ω-conotoxin</t> <t>GVIA).</t> The current measured after each toxin was subtracted from that evoked before each treatment to determine the current carried by L-, P/Q- and N-type channels. The remaining current was attributed to R-type channels. ( E ) Examples of N-, L- and WCCC elicited by voltage steps in controls and TG treated cells. ( F ) The resulting current density for each current type normalised for the peak maximal current was averaged (n = 5) to give the average current density from neurons treated with vehicle or thapsigargin. ( G ) In a subsequent experiment L- or N-type channel toxins were administered, alternating the order (n = 5 for each condition). Mean±S.E.M are shown and compared by t-test. * P <0.05 vs control.
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    Alomone Labs conotoxin gvia
    Summary of the compounds and antiepileptic drugs evaluated in the calcium oscillations assay.
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    Alomone Labs ω conotoxin gvia ω gvia
    SPC-01 generates functional neurons. (a) Preincubation with Cd 2+ (100 μ M ) together with Ni 2+ (50 μ M ), for 5 minutes significantly reduced the [Ca 2+ ] i responses induced by K + in all cells tested by 93% ± 9.2% ( n = 5), indicating the involvement of voltage-activated Ca 2+ channels in depolarization-induced Ca 2+ entry. (b) The specific L-type Ca 2+ channel blocker nicardipine (1 μ M ) reduced the [Ca 2+ ] i responses by 28% ± 7% ( n = 5; P = 0.002). The trace in (c) shows the [Ca 2+ ] i responses observed in a selected SPC-01-derived neuron induced by 50 m M K + , preincubated for 2 minutes with <t>Ω</t> <t>-GVIA</t> (800 n M ) and then stimulated with K + . After a washout of 10 minutes, the same cells were subjected to K + to observe recovery. Similarly, the effect of the P/Q-Ca 2+ channel blocker, Ω -Aga-IVA, 300 n M was tested before and during stimulation with high K + (d) . After washout of the toxin, the [Ca 2+ ] i response was shown to be reversible. The histogram (e) summarizes the results presented in (c) and (d) . The resting [Ca 2+ ] i level in these cells is indicated as basal. The results are expressed as mean ± SEM; n = 4 (Ω-GVIA) and n = 5 (Ω-Aga-IVA). Asterisks indicate the statistical significance ( P > 0.05) versus control K + stimulus.
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    Alomone Labs ω conotoxin gvia c 300
    The effect of eupatilin in combination <t>with</t> <t>ω-conotoxin</t> <t>GVIA</t> or ω-agatoxin IVA on 4-AP-induced release of glutamate in rat cerebral cortex synaptosomes. Eupatilin was added 10 min before 4-AP addition and ω-conotoxin GVIA or ω-agatoxin IVA was added 20 min before this. Data are the mean ± SEM. ***, p < 0.001, as compared to the control; #, p < 0.05, as compared to the ω-conotoxin GVIA-treated group; n = 5 per group.
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    ( A ) Isolated magnocellular neurons were identified as either vasopressin or oxytocin by their presence or absence of the GFP signal. Scale bar = 10 µm. ( B ) The current-voltage relationships show no difference between whole-cell calcium currents (WCCC) of vasopressin and oxytocin neurons (n = 20 each). ( C ) Normalized conductance (G/G max , fitted using Boltzmann equation) showed no significant change with or without TG. ( D ) Examples of two neurons (both oxytocin neurons), one vehicle treated (•) and one 50 min after pre-treating with thapsigargin (○). The steady-state Ca 2+ current is plotted against time following treatment with channel toxins (in order: 1 µM nicardipine; 200 nM ω-agatoxin IV; 500 nM ω-conotoxin GVIA). The current measured after each toxin was subtracted from that evoked before each treatment to determine the current carried by L-, P/Q- and N-type channels. The remaining current was attributed to R-type channels. ( E ) Examples of N-, L- and WCCC elicited by voltage steps in controls and TG treated cells. ( F ) The resulting current density for each current type normalised for the peak maximal current was averaged (n = 5) to give the average current density from neurons treated with vehicle or thapsigargin. ( G ) In a subsequent experiment L- or N-type channel toxins were administered, alternating the order (n = 5 for each condition). Mean±S.E.M are shown and compared by t-test. * P <0.05 vs control.

    Journal: PLoS ONE

    Article Title: The Involvement of Voltage-Operated Calcium Channels in Somato-Dendritic Oxytocin Release

    doi: 10.1371/journal.pone.0025366

    Figure Lengend Snippet: ( A ) Isolated magnocellular neurons were identified as either vasopressin or oxytocin by their presence or absence of the GFP signal. Scale bar = 10 µm. ( B ) The current-voltage relationships show no difference between whole-cell calcium currents (WCCC) of vasopressin and oxytocin neurons (n = 20 each). ( C ) Normalized conductance (G/G max , fitted using Boltzmann equation) showed no significant change with or without TG. ( D ) Examples of two neurons (both oxytocin neurons), one vehicle treated (•) and one 50 min after pre-treating with thapsigargin (○). The steady-state Ca 2+ current is plotted against time following treatment with channel toxins (in order: 1 µM nicardipine; 200 nM ω-agatoxin IV; 500 nM ω-conotoxin GVIA). The current measured after each toxin was subtracted from that evoked before each treatment to determine the current carried by L-, P/Q- and N-type channels. The remaining current was attributed to R-type channels. ( E ) Examples of N-, L- and WCCC elicited by voltage steps in controls and TG treated cells. ( F ) The resulting current density for each current type normalised for the peak maximal current was averaged (n = 5) to give the average current density from neurons treated with vehicle or thapsigargin. ( G ) In a subsequent experiment L- or N-type channel toxins were administered, alternating the order (n = 5 for each condition). Mean±S.E.M are shown and compared by t-test. * P <0.05 vs control.

    Article Snippet: The channel blockers used were: L-type, 1 µM nicardipine (Sigma-Aldrich, UK); P/Q-type, 0.2 µM ω-agatoxin IVA (Alomone Labs, Jerusalem, Israel); N-type, 0.5 µM ω-conotoxin GVIA (Alomone Labs, Jerusalem, Israel); R-type, 0.02 µM SNX 482 (Alomone Labs, Jerusalem, Israel).

    Techniques: Isolation

    Summary of the compounds and antiepileptic drugs evaluated in the calcium oscillations assay.

    Journal: PLoS ONE

    Article Title: New In Vitro Phenotypic Assay for Epilepsy: Fluorescent Measurement of Synchronized Neuronal Calcium Oscillations

    doi: 10.1371/journal.pone.0084755

    Figure Lengend Snippet: Summary of the compounds and antiepileptic drugs evaluated in the calcium oscillations assay.

    Article Snippet: Diazepam was from Fragon; phenytoin from Fluka; conotoxin GVIA and TTX from Alomone Labs; gabapentin from TCI Chemicals; and levetiracetam and retigabine were synthetized at UCB.

    Techniques:

    SPC-01 generates functional neurons. (a) Preincubation with Cd 2+ (100 μ M ) together with Ni 2+ (50 μ M ), for 5 minutes significantly reduced the [Ca 2+ ] i responses induced by K + in all cells tested by 93% ± 9.2% ( n = 5), indicating the involvement of voltage-activated Ca 2+ channels in depolarization-induced Ca 2+ entry. (b) The specific L-type Ca 2+ channel blocker nicardipine (1 μ M ) reduced the [Ca 2+ ] i responses by 28% ± 7% ( n = 5; P = 0.002). The trace in (c) shows the [Ca 2+ ] i responses observed in a selected SPC-01-derived neuron induced by 50 m M K + , preincubated for 2 minutes with Ω -GVIA (800 n M ) and then stimulated with K + . After a washout of 10 minutes, the same cells were subjected to K + to observe recovery. Similarly, the effect of the P/Q-Ca 2+ channel blocker, Ω -Aga-IVA, 300 n M was tested before and during stimulation with high K + (d) . After washout of the toxin, the [Ca 2+ ] i response was shown to be reversible. The histogram (e) summarizes the results presented in (c) and (d) . The resting [Ca 2+ ] i level in these cells is indicated as basal. The results are expressed as mean ± SEM; n = 4 (Ω-GVIA) and n = 5 (Ω-Aga-IVA). Asterisks indicate the statistical significance ( P > 0.05) versus control K + stimulus.

    Journal: Stem Cell Research & Therapy

    Article Title: Conditionally immortalized stem cell lines from human spinal cord retain regional identity and generate functional V2a interneurons and motorneurons

    doi: 10.1186/scrt220

    Figure Lengend Snippet: SPC-01 generates functional neurons. (a) Preincubation with Cd 2+ (100 μ M ) together with Ni 2+ (50 μ M ), for 5 minutes significantly reduced the [Ca 2+ ] i responses induced by K + in all cells tested by 93% ± 9.2% ( n = 5), indicating the involvement of voltage-activated Ca 2+ channels in depolarization-induced Ca 2+ entry. (b) The specific L-type Ca 2+ channel blocker nicardipine (1 μ M ) reduced the [Ca 2+ ] i responses by 28% ± 7% ( n = 5; P = 0.002). The trace in (c) shows the [Ca 2+ ] i responses observed in a selected SPC-01-derived neuron induced by 50 m M K + , preincubated for 2 minutes with Ω -GVIA (800 n M ) and then stimulated with K + . After a washout of 10 minutes, the same cells were subjected to K + to observe recovery. Similarly, the effect of the P/Q-Ca 2+ channel blocker, Ω -Aga-IVA, 300 n M was tested before and during stimulation with high K + (d) . After washout of the toxin, the [Ca 2+ ] i response was shown to be reversible. The histogram (e) summarizes the results presented in (c) and (d) . The resting [Ca 2+ ] i level in these cells is indicated as basal. The results are expressed as mean ± SEM; n = 4 (Ω-GVIA) and n = 5 (Ω-Aga-IVA). Asterisks indicate the statistical significance ( P > 0.05) versus control K + stimulus.

    Article Snippet: Sandimmune (Novartis Pharma AG, Basel, Switzerland); Immuran (GlaxoSmith-Kline); Solu-Medrol (Pfizer, Puurs, Belgium); Fura-2 AM 1 m M solution in anhydrous DMSO and Pluronic F-127 (30% stock in distilled water) (Molecular Probes, USA); ω-Aga Toxin IVA (ω-Aga IVA), and ω-conotoxin GVIA (ω-GVIA) (Alomone Labs Ltd. Jerusalem, Israel).

    Techniques: Functional Assay, Derivative Assay

    The effect of eupatilin in combination with ω-conotoxin GVIA or ω-agatoxin IVA on 4-AP-induced release of glutamate in rat cerebral cortex synaptosomes. Eupatilin was added 10 min before 4-AP addition and ω-conotoxin GVIA or ω-agatoxin IVA was added 20 min before this. Data are the mean ± SEM. ***, p < 0.001, as compared to the control; #, p < 0.05, as compared to the ω-conotoxin GVIA-treated group; n = 5 per group.

    Journal: International Journal of Molecular Sciences

    Article Title: Inhibition of Synaptic Glutamate Exocytosis and Prevention of Glutamate Neurotoxicity by Eupatilin from Artemisia argyi in the Rat Cortex

    doi: 10.3390/ijms232113406

    Figure Lengend Snippet: The effect of eupatilin in combination with ω-conotoxin GVIA or ω-agatoxin IVA on 4-AP-induced release of glutamate in rat cerebral cortex synaptosomes. Eupatilin was added 10 min before 4-AP addition and ω-conotoxin GVIA or ω-agatoxin IVA was added 20 min before this. Data are the mean ± SEM. ***, p < 0.001, as compared to the control; #, p < 0.05, as compared to the ω-conotoxin GVIA-treated group; n = 5 per group.

    Article Snippet: Fura-2-acetoxymethyl ester (Fura-2-AM), DiSC 3 (5), and antibodies against glutaminase (#701965), p-DAPK1 (Ser736; PA5-105872) were acquired from Invitrogen (Carlsbad, CA, USA). ω-conotoxin GVIA (C-300) and ω-agatoxin IVA (STA-500) were acquired from Alomone (Jerusalem, Israel).

    Techniques: