Fig. 4: Insular Gi-DREADD activation decreases BNST fos expression after restraint stress, and insular transients are associated with struggling bouts. | Nature Communications

Fig. 4: Insular Gi-DREADD activation decreases BNST fos expression after restraint stress, and insular transients are associated with struggling bouts.

From: Delineation of an insula-BNST circuit engaged by struggling behavior that regulates avoidance in mice

Fig. 4: Insular Gi-DREADD activation decreases BNST fos expression after restraint stress, and insular transients are associated with struggling bouts.The alternative text for this image may have been generated using AI.

a Timeline. Mice were injected with a Gi-DREADD (hM4Di) coding virus in one hemisphere (red) and an eGFP control in the other (cyan). Three weeks later, mice were injected with CNO (3 mg/kg i.p.). After 30 min, they were restrained for 1 h. Animals were processed 30 min later for fluorescent in situ hybridization. b Example fluorescent in situ hybridization from the BNST with examples of crh only (magenta box), fos only (cyan box), and crh/fos doubly labeled (white box) cells. c Comparison of restraint stress (burgundy) vs. nonstress (teal) and/or CNO (gray) or saline (blue) exposure on percent of fos-positive crh cells in the BNST ipsilateral to the Gi-DREADD injection (darker shading) vs. the BNST ipsilateral to the eGFP control (lighter shading). Paired t test (two-sided), t(5) = 7.122, ***P = 0.0008. d Schematic. Virus coding for either GCaMP7f (n = 9), SF-iGluSnFR (n = 10), or iGABASnFR (n = 5) was injected into the insula, followed by fiber placement. e Bout-associated signals from GCaMP (purple), SF-iGluSnFR (blue), or iGABASnFR (yellow) across six 30-min restraint exposures (see Fig. 2g). Lighter shading: s.e.m. across bouts. f Bout-associated signals by animal. g Correlation coefficient (Pearson, no adjustments for multiple comparisons) of the relationship between AUC from 0 to 5 s and bout length over days. GCaMP (purple circle), SF-iGluSnFR (blue triangle), and iGABASnFR (yellow square). Correlation coefficient shape is solid if P < 0.05 and open if P ≥ 0.05. Rimonabant was administered to GCaMP and iGABASnFR on day 5. SF-iGluSnFR experiment finished on day 4. h–j Slope of AUC vs. bout length by day with 95% confidence interval. h GCaMP (purple). i SF-iGluSnFR (blue). j iGABASnFR (yellow). k Percent of GCaMP transient number normalized to day 1 (set at 100%). (n = 9) Error bars: s.e.m. across animals. Colors: saline (blue) and rimonabant (peach). l Frequency of GCaMP transients during restraint on day 1 (30 min after i.p. saline injection) vs. day 5 (30 min after i.p. rimonabant injection). Paired t test (two-sided), t(8) = −5.4881, ***P = 0.0006. m Frequency of GCaMP transients during restraint on day 4 (30 min after i.p. saline injection) vs. day 5 (30 min after i.p. rimonabant injection). Paired t test (two-sided), t(8) = −2.3889, *P = 0.043. l, m Error bars: s.e.m. across bouts (n = 9).

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