Fig. 4: Chemogenetic inhibition of CeAL CRF + neurons reduced CeAL c-Fos + expression and, in males, reduced activity of SOM + cells.
a Experimental design. b Representative image of CeAL labeled with DAPI, c-Fos +, and SOM + . c CeAL CRF + neuron inhibition during startle testing reduced c-Fos + expression in CeAL (Unpaired, Two-Tailed: t = 2.26, df = 43, p = 0.02, d = 0.67). n = 23 (mCherry: mCh), 22 (hM4di). d Suppressed c-Fos+ as a result of CeAL CRF + neuron inhibition occurred independent of rearing (3-way-ANOVA virus F(1, 37) = 4.26, p = 0.04, η2 = 0.33; rearing F(1, 37) = 1.91, p = 0.17, η2 = 0.27). n = 7 (mCh M Ctrl), n = 6 (mCh F Ctrl, hM4Di F Ctrl, mCh F ELA, hM4Di F ELA, hM4Di M Ctrl), n = 4 (mCh M ELA, hM4Di M ELA). e CeAL CRF + neuron inhibition during startle testing reduced c-Fos + co-expression with SOM + in CeAL (Unpaired, Two-Tailed: t = 2.83, df = 43, p = 0.007, d = 1.25). n = 23 (mCh), 22 (hM4di). f Reduced co-expression of c-Fos + with SOM+ following CeAL CRF+ neuron inhibition was sex-dependent (3-way-ANOVA virus x sex F(1, 37) = 15.82, p = 0.0003, η2 = 0.65), occurring in male Ctrl (mCh vs Di: p = 0.001) and male ELA (mCh vs Di: p = 0.01) mice but not female Ctrl (mCh vs Di: p > 0.99) or female ELA (mCh vs Di: p > 0.99) mice. Further, male mice expressing mCherry virus showed greater co-expression compared to female mCherry-expressing mice (M Ctrl vs F Ctrl: p = 0.002; M ELA vs F ELA: p = 0.03). Lastly, ELA reared mice exhibited greater co-expression compared to Ctrls (3-way-ANOVA rearing F(1, 37) = 15.47, p = 0.004, η2 = 0.64). n = 7 (mCh M Ctrl), n = 6 (mCh F Ctrl, hM4Di F Ctrl, mCh F ELA, hM4Di F ELA, hM4Di M Ctrl), n = 4 (mCh M ELA, hM4Di M ELA). All plots depict individual mice and means ± SEM. *< 0.05, **< 0.01, ***< 0.001. Source data are provided as a Source Data file. Figure 4a created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.
