Fig. 8: Optogenetic manipulations of input from avTRN drive rebound responses in NAc-projecting PVT neurons. | Nature Communications

Fig. 8: Optogenetic manipulations of input from avTRN drive rebound responses in NAc-projecting PVT neurons.

From: Convergent direct and indirect cortical streams shape avoidance decisions in mice via the midline thalamus

Fig. 8

a, b Schematic of the experimental approach for fiber photometry imaging of NAc-projecting PVT cells while optogenetically inhibiting avTRN-PVT pathway, and fiber placement around pPVT (n = 3 mice per group). c, d Heatmaps of calcium responses for mCherry and Halo subjects (left two) and average calcium signal (right) before 2AA. e Calcium signal AUC quantification for mCherry and Halo subjects, two-way ANOVA interaction provided (n = 30 Trials from 3 mice for each group); F(2, 116) = 0.08, P = 0.92. f, g Heatmaps (f) and averages (g) of calcium responses for mCherry and Halo subjects from pre 5-s to post 5-s of light onset from (c). Mixed-effects model (REML) interaction reported in graph. h, i Adapted from Fig. 7b, during 2AA. Heatmaps (h) and averages (i) of calcium responses for Light-off and Light-on sessions from 10-s prior to WS onset. Mixed-effects model (REML) interaction reported in graphs, Avoidance: (Off, n = 205 Trials; On, n = 192 Trials); Escape: P < 0.001 (Off, n = 35 Trials; On, n = 48 Trials). j Top: Schematic of the experimental approach for fiber photometry imaging of NAc-projecting PVT cells and optogentically activating avTRN-PVT pathway. Bottom: Representative images of GCaMP7s and ChrimsonR-tdTomato expression, and fiber placement around pPVT. k Heatmap of calcium responses during optical stimulation. l Average calcium signal. m Calcium signal AUC quantification for each stage. One-way ANOVA interaction; F(2, 87) = 144.2, P < 0.0001 (n = 30 Trials from 3 mice). n Left: Schematic of the experimental approach for ex vivo single-cell slice imaging of GCaMP during TRN optogenetic activation. Right: Example 4x image of slice with GCaMP8f expression. o Averaged z-score calcium signal before and after bath application of PTX (50 µM) and CGP55845 (5 µM) during optogenetic stimulation of TRN normalized to the slope of baseline. p Calcium signal AUC quantification before and after bath application of GABA antagonists. Two-way repeated measures ANOVA interaction, F(2, 266) = 31.74, P < 0.0001 (n = 134 cells from 5 mice). For all quantifications, multiple comparisons were conducted corrected by two-stage linear step-up procedure, black lines along x-axis indicate significant changes reported between groups. Asterisks indicate where ANOVA multiple comparisons found significance. Data are shown as mean ± s.e.m. Source data are provided as a Source Data file.

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