anti cav3 1  (Thermo Fisher)

Journal: Nature Communications

Article Title: Thalamocortical bursts encode reward contingencies and drive associative learning

doi: 10.1038/s41467-026-76581-6

Figure Lengend Snippet: a Schematic of shRNA viral injection to knock down (KD) Ca V 3.1 in the VPM and POm (left panel), including a scrambled shRNA construct used as a control to account for non-specific effects of viral expression. Representative histological image (middle panel) showing shRNA expression (yellow, shRNA-mOrange) in the thalamus with DAPI counterstain (blue). Higher magnification insets (right panels) show mOrange fluorescence (yellow) and Ca V 3.1 expression (via anti-Ca V 3.1 immunofluorescence) in KD conditions. Each condition included n = 5 animals; histological quantification was performed on n = 18 brain slices from 2 scrambled shRNA control animals and n = 34 brain slices from 4 Ca V 3.1 KD animals. b Proportion of shRNA expression within anatomical boundaries of VPM and POm ( n = 34 brain slices from 4 animals). c Normalized thalamic anti-Ca V 3.1 fluorescence intensity of Ca V 3.1 KD animals ( n = 34 brain slices from 4 animals) compared to scrambled shRNA controls ( n = 18 brain slices from 2 animals) ( p = 1.1514 × 10⁻⁴). d Trial-by-trial learning curves in Ca V 3.1 KD mice compared to scrambled shRNA controls (left panel). Data is shown as cohort mean with SEM as shaded areas. Learning speed quantified as the number of trials to reach expert performance in Ca V 3.1 KD animals and scrambled shRNA controls (right panel, n = 5 mice in each group, p = 0.0317). e Schematic of systemic Z944 or DMSO (control) i.p. injection in mice. f Session-wise performance (d′) in expert animals (dashed grey line) following systemic administration of Z944 (red) compared to DMSO control sessions (grey). Lines represent the cohort means with SEM as shaded areas (4 sessions from n = 6 animals). g Same as in d for Z944-treated animals and DMSO controls ( n = 4 mice in each group, p = 0.0286). h Example reconstruction of a 4-shank Neuropixel recording in POm and VPM. i Overall burstiness (see methods) during 15 min recordings in control-DMSO, Z944 and Ca V 3.1 KD conditions. VPM and POm units were pooled, DMSO and Z944 n = 422 (same units), shRNA, n = 305 units. Statistical comparison was performed using a Kruskal–Wallis test (H(2) = 111.22, p < 0.001). j Top: raster plots of example POm units during DMSO and Z944 (left, middle, same unit) and Ca V 3.1 KD (right). Middle: Burst firing rate for wide (rewarded) and narrow (unrewarded) aperture relative to whisker touch (dashed line). Significant differences ( α = 0.05) are indicated with dashed lines. Only touch-modulated POm and VPM units were selected (same units in DMSO and Z944 conditions, n = 140, Z944, n = 140, shRNA, n = 51). Bottom: Distribution of mean burst indices for the same units as above. Statistical comparisons performed using two-tailed Wilcoxon signed-rank tests: DMSO, p = 1.1 × 10⁻⁵; Z944, p = 0.0506; shRNA, p = 0.223. n.s.: p > 0.05, *: p ≤ 0.05, **: p ≤ 0.01 and ***: p ≤ 0.001; c , d , g : two-tailed Mann–Whitney U -Test, f: two-tailed, paired t-test, i: Kruskal-Wallis with post-tests. j : two-tailed Welch’s t -test with Benjamini-Hochberg correction for multiple comparisons (middle panels); two-tailed Wilcoxon signed-rank test (lower panels). For detailed description of statistics see Supplementary Data . Source data are provided as a file.

Article Snippet: Sections containing the thalamus were processed for immunohistochemistry using an antibody against the Ca V 3.1 channel (Anti-CACNA1G (Ca V 3.1) Antibody, #ACC-021, Alomone Labs, Jerusalem 9104201, Israel), diluted 1: 200 in phosphate buffer, and counterstained with DAPI to visualize cytoarchitecture.

Techniques: shRNA, Injection, Knockdown, Construct, Control, Expressing, Fluorescence, Immunofluorescence, Comparison, Whisker Assay, Two Tailed Test, MANN-WHITNEY