snx482  (Alomone Labs)

Journal: Neuroscience

Article Title: Regulation of glutamatergic and GABAergic neurotransmission in the chick nucleus laminaris: role of N-type calcium channels

doi: 10.1016/j.neuroscience.2009.09.013

Figure Lengend Snippet: Glutamatergic transmission in the NL is predominantly triggered by Ca 2+ entry through N-type VGCCs. A : Both the ipsilateral (open circles) and the contralateral (filled circles) EPSCs recorded from the same NL neuron were largely and irreversibly blocked by the N-type blocker ω-Conotoxin-GVIA (ω-CTx-GVIA, 2.5 μM), with nearly identical time courses. Averaged traces under control (thin traces) and drug (thick traces) conditions are shown on the right. Stimulus artifacts are truncated for clarity. B-C : No inhibition of EPSCs was produced by the P/Q-type blocker ω-Agatoxin-IVA (ω-Aga-IVA, 0.1 μM) or the L-type blocker nimodipine (10 μM). D: A small and reversible inhibition of the EPSCs by the R-type blocker SNX-482 (50 nM) was observed. The different time courses of the EPSCs observed in the sampled neurons may be due to different locations of the neurons along the tonotopic frequency axis. E : Summary of the effects of different VGCC blockers on EPSCs of NL neurons. ω-CTx-GVIA inhibited the ipsilateral and the contralateral EPSCs by 88% and 86%, respectively. Blockers for P/Q- and L-type VGCCs had no effects on EPSCs. SNX-482 produced a small inhibition with a large variation among cells. No differences in the percent inhibition by either drug were detected between the ipsilateral and the contralateral EPSCs. Error bars represent standard deviations. n.s.: non-significant (paired t-test, p > 0.05; the number of cells is indicated in parentheses).

Article Snippet: All chemicals and drugs were obtained from Sigma (Saint Louis, MO), except for SNX-482, ω-Conotoxin-GVIA (ω-CTx-GVIA) and tACPD, which were obtained from Peptide International (Louisville, KY), Alomone Labs (Jerusalem, Israel), and Tocris (Ballwin, MO), respectively.

Techniques: Transmission Assay, Inhibition, Produced

Journal: bioRxiv

Article Title: Alternative splicing of auxiliary β2-subunits stabilizes Cav2.3 Ca2+ channel activity in continuously active midbrain dopamine neurons

doi: 10.1101/2021.02.10.430224

Figure Lengend Snippet: SNX-482 inhibition of non-L-type I Ca in cultured midbrain DA neurons. A. Representative traces illustrating the inhibition of non-L-type I Ca by 100 nM SNX-482 (red). Cells were initially perfused with a bath solution containing 3 μM isradipine (ISR, black). Full block was obtained using 2 μM Cd 2+ (blue). Square pulses (50 ms) were applied to 0 mV from a holding potential of −70 mV (top) B. Current amplitude values plotted as a function of time. After stabilization of I Ca with ISR (black circles), 100 nM SNX-482 was applied. The remaining currents was blocked by 2 μM Cd 2+ . C. SNX-482 inhibition expressed as % of control I Ca after LTCC block using 3 μM ISR. D. Mean current amplitude at the end of ISR application and at the end of SNX-482 application. Data represent the means ± SEM for the indicated number of experiments (N=4). Statistical significance was determined using paired Student’s t-test: *** p

Article Snippet: For inhibition experiments, 100 nM SNX-482 (Alomone, Cat # RTS-500 dissolved in ACSF) or 10 µM nifedipine (Alomone, Cat # N-120 diluted into ACSF from a freshly prepared 10 mM stock solution in DMSO) was bath applied (in ACSF).

Techniques: Inhibition, Cell Culture, Blocking Assay

Journal: bioRxiv

Article Title: Alternative splicing of auxiliary β2-subunits stabilizes Cav2.3 Ca2+ channel activity in continuously active midbrain dopamine neurons

doi: 10.1101/2021.02.10.430224

Figure Lengend Snippet: SNX-482 effects on pacemaking of cultured mouse midbrain DA neurons. A. Representative recording of spontaneous firing activity of cultured midbrain dopaminergic neurons before, during and after the application (wash-out) of 100 nM SNX-482. Inset (bottom right): overlay of one single AP before (control) and during the application of 100 nM SNX-482. B. Firing frequency [Hz], coefficient of variation of the interspike interval [%], and AHP peak [mV] before (control) and during the application of 100 nM SNX-482. Data represent the means ± SEM for the indicated number of experiments (N=3). Statistical significance was determined using paired Student’s t-test.: *** p

Article Snippet: For inhibition experiments, 100 nM SNX-482 (Alomone, Cat # RTS-500 dissolved in ACSF) or 10 µM nifedipine (Alomone, Cat # N-120 diluted into ACSF from a freshly prepared 10 mM stock solution in DMSO) was bath applied (in ACSF).

Techniques: Cell Culture, Activity Assay

Journal: Journal of Neurophysiology

Article Title: Low-threshold calcium currents contribute to locomotor-like activity in neonatal mice

doi: 10.1152/jn.00583.2011

Figure Lengend Snippet: Application of the selective R-type calcium channel blocker SNX-482 does not alter fictive locomotor-like activity. A : paired extracellular recordings from iL2 and iL5 in an isolated whole-cord preparation with cocktail (3–10 μM NMDA,

Article Snippet: The cycle frequency was not significantly affected by SNX-482 (control, mean = 0.24 ± 0.04 Hz; SNX-482, mean = 0.23 ± 0.02 Hz; n = 3; t = 0.39; P = 0.72; ).

Techniques: Activity Assay, Isolation

Journal: PLoS ONE

Article Title: P2X6 Knockout Mice Exhibit Normal Electrolyte Homeostasis

doi: 10.1371/journal.pone.0156803

Figure Lengend Snippet: Compensatory mechanisms for the loss of P2x6 function in the heart. a-h) The mRNA expression levels of P2x1 (a), P2x2 (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), Trpm7 (g), Cnnm2 (h), in heart of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data represent mean (n = 10) ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice. * P

Article Snippet: Samples were then subjected to SDS-PAGE and Western blots were incubated with a rabbit anti-mouse P2x2 antibody (Alomone Labs, Jerusalem, Israel, 1:500) or a rabbit anti-mouse P2x4 (H-40, Santa Cruz biotechnology, Santa Cruz CA, USA, 1:500) in 5% milk o/n at 4°C.

Techniques: Expressing, Mouse Assay, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: P2X6 Knockout Mice Exhibit Normal Electrolyte Homeostasis

doi: 10.1371/journal.pone.0156803

Figure Lengend Snippet: Protein abundance of P2x2 and P2x4 in response to the loss of P2x6. a) Western blots of membrane and cytosol fractions of P2x6 -/- (KO) and P2x6 +/+ (WT) mice. The upper blot shows the immune-staining for P2x2 in mouse kidney material. To the right a western blot of HEK293 cells transiently transfected with HA-tagged P2x4 (P4) and mock (M) constructs is displayed. Middle, two western blots below are immune-stained for P2x4, left depicts P2x6 -/- (KO) and P2x6 +/+ (WT) material stained for P2x4, right represents a P2x4 blot on HEK293 material transiently transfected with human P2X4 and a mock construct. Bottom, displays a ß-actin immune-staining used as a loading control. Ladders (ez-run prestained marker (ThermoScientific, Breda, The Netherlands) represent protein size in kilo Dalton (kD). b) Protein expression levels for the P2x2 membrane lysate, P2x4 membrane lysate, P2x4 cytosol lysate, ß-actin membrane and ß-actin cytosol lysates in P2x6 -/- and P2x6 +/+ mice. Data (n = 3) represents mean ± SEM and are expressed as the % of total band intensity.

Article Snippet: Samples were then subjected to SDS-PAGE and Western blots were incubated with a rabbit anti-mouse P2x2 antibody (Alomone Labs, Jerusalem, Israel, 1:500) or a rabbit anti-mouse P2x4 (H-40, Santa Cruz biotechnology, Santa Cruz CA, USA, 1:500) in 5% milk o/n at 4°C.

Techniques: Western Blot, Mouse Assay, Staining, Transfection, Construct, Marker, Expressing

Journal: PLoS ONE

Article Title: P2X6 Knockout Mice Exhibit Normal Electrolyte Homeostasis

doi: 10.1371/journal.pone.0156803

Figure Lengend Snippet: P2x subunit expression in response to the loss of P2x6 function in the kidney. a-f) The mRNA expression levels of P2x1 (a), P2x2 (b), P2x3 (c), P2x4 (d), P2x5 (e), P2x7 (f), in kidney of P2x6 +/+ (Black bars), P2x6 +/- (Striped bars), P2x6 -/- (white bars) mice were measured by quantitative RT-qPCR and normalized for Gapdh expression. Data (n = 10) represent mean ± SEM and are expressed as the fold difference when compared to the expression in P2x6 +/+ mice.

Article Snippet: Samples were then subjected to SDS-PAGE and Western blots were incubated with a rabbit anti-mouse P2x2 antibody (Alomone Labs, Jerusalem, Israel, 1:500) or a rabbit anti-mouse P2x4 (H-40, Santa Cruz biotechnology, Santa Cruz CA, USA, 1:500) in 5% milk o/n at 4°C.

Techniques: Expressing, Mouse Assay, Quantitative RT-PCR