Fig. 6: The avTRN–PVT circuit controls active avoidance behavior.

a Top: Schematic of the experimental approach for fiber photometry imaging of the axon terminals of GAD2⁺ TRN cells in the pPVT. Bottom: Representative images of GCaMP7s expression and fiber placement around pPVT. b Top: Heatmaps of calcium responses for avoidance and escape trials. Bottom: Average calcium signal and WS duration, n = 5 mice. c, d Left: Average calcium responses for all avoidance (blue) and escape (red) trials during WS onset (c) and shuttle initiate events (d). Mixed-effects model (REML) interaction reported in graphs. WS onset: Avoidance, n = 214 Events; Escape, n = 86 events. Shuttle initiate: Avoidance, n = 158 events; Escape, n = 64 events. Right: Calcium signal AUC quantification in 1 s bins. WS onset REML interaction, F_(3,894) = 0.28, P = 0.84; Shuttle Initiate REML interaction F_(9,1980) = 9.19, P < 0.0001. e Top: Viral strategy schematic for optogenetic stimulation of avTRN–PVT circuit. Bottom: Representative image of mCherry-opsin expression and fiber placement around pPVT. f 2AA task schematic. g Group behavioral data across test sessions normalized to first light-off session for avoidance rates (left), latencies to avoid (middle), and latencies to escape (right). Two-way ANOVA interactions provided, (Ctl, n = 7 mice; ChR, n = 9 mice). Avoidance rate: F_{(2, 28)} = 4.73, P = 0.017, Latency to avoid: F_{(2, 28)} = 0.13, P = 0.88, Latency to escape: F_{(2, 42)} = 0.94, P = 0.4, h 2AA task schematic. i Group behavioral data across test sessions normalized to first light-off session for avoidance rates (left), latencies to avoid (middle), and latencies to escape (right). Two-way ANOVA interactions provided (Ctl, n = 6 mice; ChR, n = 7 mice). Avoidance rate: F_{(2, 22)} = 4.09, P = 0.031. Latency to avoid: F_{(2, 21)} = 5.96, P = 0.0089. Latency to escape: F_{(2, 22)} = 0.39, P = 0.68. 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, and red or blue lines denote the first significant change from the previous bin for within trial type comparisons. 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.