Journal: PLoS ONE

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

doi: 10.1371/journal.pone.0042602

Figure Lengend Snippet: A , The current density versus test potential ( I–V ) relationship for R- and S- I Na . B , Dot plots represent the membrane capacitance of DRG neurons displaying R- (open circles) and S- I Na (filled circles). Vertical bars represent the median. *** P<0.001, Mann-Whitney U -test. C , Concentration-response curves for block of the R- and S- I Na by tetrodotoxin (TTX). Solid lines represent nonlinear regression least-square fits of experimental points to a Hill equation (see under ). D , The voltage-dependence of the Na + conductance ( G Na , left panel) and steady-state inactivation (right panel) of R- and S- I Na . Conductance was calculated from G Na = I Na /(V m −V rev ) , in which I Na is the peak current, V m is the potential of the test pulse, and V rev is the reversal potential for I Na . The solid line represents a nonlinear regression fit to the Boltzmann function: 1/(1+exp[−( V − V 1/2 )/ k ]), where V is the step membrane potential, V 1/2 is the half-activation potential, and k is a slope factor. Voltage-dependence of inactivation was determined using a 200 ms conditioning pulse followed by a test pulse to −10 mV. Test pulse currents were normalized to the maximal value. Solid line is a fit to the Boltzmann equation ( k is negative for inactivation curve). Error bars on each symbol represent the mean ± s . e . m . The number of neurons tested is shown in parentheses.

Article Snippet: Stock solutions were made for the following drugs: γ-aminobutyric acid (GABA), l -glutamic acid hydrochloride, oxotremorine M, FPL64176 (all from TOCRIS Cookson Ltd., Ellisville, MO), nifedipine (EMD chemicals, Gibbstown, NJ), tetrodotoxin (TTX), ω-agatoxin IVA, ω-conotoxin GVIA, SNX-482 (all from Alomone Labs, Jerusalem, Israel), (±)-norepinephrine, serotonin hydrochloride (5-hydroxytryptamine, 5-HT) and CdCl 2 (all from Sigma-Aldrich).

Techniques: Membrane, MANN-WHITNEY, Concentration Assay, Blocking Assay, Activation Assay

Journal: Frontiers in Immunology

Article Title: The Therapeutic Effect of STAT3 Signaling-Suppressed MSC on Pain and Articular Cartilage Damage in a Rat Model of Monosodium Iodoacetate-Induced Osteoarthritis

doi: 10.3389/fimmu.2018.02881

Figure Lengend Snippet: Effects of STAT3 inhibition on anti-inflammatory activity and migration potential in MSCs from osteoarthritis (OA) patients. (A) OA-MSCs were incubated with STA21 for 3 days and, ELISA was performed to measure IL-6, IL-8, IL-1β, and VEGF concentrations. (B) OA-MSCs were incubated with STA21 for 1 h. The expression of phospho (p)-STAT3 705, 727, and total STAT3 protein was analyzed by Western blotting. (C) mRNA levels for anti-inflammatory cytokines IL-10 and TGF-β were measured by real time-PCR (D) Expression of chemokine receptors in OA-MSCs incubated with STA21 was measured by real time-PCR. OA-MSC were isolated of fat tissues obtain from each 3 person with OA. Data represent the mean ± SD of 3 independent experiments ( * P < 0.05; ** p < 0.01).

Article Snippet: The cells were cultured for 4 days until 90% confluent (passage 0) and were then expanded for 2–3 passages and used for experiments. iSTAT3 OA-MSCs were prepared by treatment of OA-MSCs with 10 μM STA21 (Santa Cruz, Texas, USA) for 72 h.

Techniques: Inhibition, Activity Assay, Migration, Incubation, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Isolation

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

Article Title: The ducky 2J mutation in Cacna2d2 results in reduced spontaneous Purkinje cell activity and altered gene expression

doi: 10.1523/JNEUROSCI.3080-06.2006

Figure Lengend Snippet: A: Example of the effect of ω-agatoxin-IVB on the time-course of instantaneous spontaneous firing frequency of a +/+ PC, recorded in cell-attached mode at 34°C and with synaptic blockers in the control solution. The cell fires tonically in control. In the presence of BSA (25 μg/ml), the firing rate increases slightly for about 5 min. When ω-agatoxin-IVB (100 nM) is added to the superfusate, a dramatic increase in the frequency is initially observed, which develops into an irregular bursting firing pattern. The cell then enters a quiescent state for the rest of the recording in cell-attached mode (about 5 min). 2 s blocks of data are shown above the graph for the periods indicated by the arrows. Similar data were obtained in three other cells that were initially firing tonically.

Article Snippet: Data are given as mean ± s.e.m. and statistical significances between groups were determined by Student’s unpaired t test, or by analysis of variance (ANOVA) followed by Tukey’s multiple comparison test, or using a non-parametric test, as appropriate Drugs used were: ω-agatoxin-IVA and-IVB (Alomone Labs, Jerusalem, Israel), which were applied in bovine serum albumin (BSA, 25 μg/ml), penitrem A and apamin (Alomone).

Techniques:

Journal: The Journal of General Physiology

Article Title: Arrangement of Kv1 α subunits dictates sensitivity to tetraethylammonium

doi: 10.1085/jgp.200910398

Figure Lengend Snippet: Pharmacological properties of homomeric and heteromeric Kv1 channels

Article Snippet: Kaliotoxin (KTX), agitoxin-1 (AgTX1), and tityustoxin-Kα (TsTX-Kα) were purchased from Alomone Labs.

Techniques:

Journal: The Journal of General Physiology

Article Title: Arrangement of Kv1 α subunits dictates sensitivity to tetraethylammonium

doi: 10.1085/jgp.200910398

Figure Lengend Snippet: Adjacently and diagonally arranged Kv1.1 and 1.2 tandem-linked gene constructs gave channels that were distinguished by TEA, but not AgTX1, TsTX-Kα, or KTX. (A) Current traces recorded using QPatch from an adjacent (Kv1.1-1.1-1.2-1.2) and diagonal (Kv1.2-1.1-1.2-1.1) channel in the absence (black) and presence of TEA, AgTX1, TsTX-Kα, or KTX (blue). (B) Dose–response curves for the forward and reverse adjacent channels show an ∼10-fold lower affinity for TEA than the corresponding diagonals. ○, Kv1.1-1.1-1.2-1.2; n = 5 cells for each concentration, manual patch clamp; Hill equation fit IC 50 = 9.6 mM, slope = 0.8. •, Kv1.2-1.2-1.1-1.1; Qpatch, n = 2–3; 8.2 mM, 0.6. □, Kv1.1-1.2-1.1-1.2; n = 4, Qpatch; 0.9 mM, 0.6. (blue) □, Kv1.1-1.2-1.1-1.2; n = 6–8, manual patch clamp; 1.1 mM, 0.6. ▪, Kv1.2-1.1-1.2-1.1; n = 3–5, Qpatch; 0.8 mM, 0.7. (Inset) Non-concatenated (red ▴; n = 6–7; 0.4 mM, 0.6) and concatenated (black ▴; n = 2–4; 0.7 mM, 0.7) homotetrameric Kv1.1 channels showed almost identical sensitivity to TEA, as with the less sensitive homotetrameric Kv1.2 (red ▾; n = 2–4; 41 mM, 0.6) channel and the tandem-linked homotetramer (black ▾; n = 3; 36 mM, 0.5). All homomer data were from Qpatch. (C) Channel dimers showed TEA susceptibilities similar to that for the pair of adjacently arranged tetrameric concatamers. □, Kv1.1-1.2; n = 8; 9.3 mM, 1.0. •, Kv1.2-1.1; n = 5–6; 7.8 mM, 0.7. ◆, Kv1.1-1.2 + Kv1.2-1.1; n = 9–11; 9.5 mM, 0.4. Data were from Qpatch. Error bars represent ± SEM.

Article Snippet: Kaliotoxin (KTX), agitoxin-1 (AgTX1), and tityustoxin-Kα (TsTX-Kα) were purchased from Alomone Labs.

Techniques: Construct, Concentration Assay, Patch Clamp