Fig. 1: ELA rearing enhanced startle and resulted in the preferential recruitment of CeAL CRF + neurons in female ELA mice. | Nature Communications

Fig. 1: ELA rearing enhanced startle and resulted in the preferential recruitment of CeAL CRF + neurons in female ELA mice.

From: Central amygdala CRF+ neurons promote heightened threat reactivity following early life adversity in mice

Fig. 1

a Experimental design. Female (F), male (M), control (Ctrl), and ELA (ELA) reared mice underwent the startle task in adulthood. b Startle in Block 1 was enhanced by ELA and sex (2-way-ANOVA rearing F(1, 53) = 16.34, p = 0.0002, η2 = 0.55; 2-way-ANOVA sex F(1, 53) = 6.92, p = 0.01, η2 = 0.36) and not an interaction (2-way-ANOVA sex x rearing F(1,53) = 0.98, p = 0.32, η2 = 0.13). n = 13 (F Ctrl), 16 (F ELA), 15 (M Ctrl), 16 (M ELA). These data have been replicated in three independent experiments. c Startle was greater in CS + trials and was enhanced by ELA (3-way-RM-ANOVA cue F(1, 53) = 50.67, p = 2.9e-9, η2 = 0.97; 3-way-RM-ANOVA rearing F(1, 53) = 15.86, p = 0.0002, η2 = 0.87). Startle did not differ by sex or an interaction (3-way-ANOVA sex F(1,53 = 2.78, p = 0.10, η2 = 0.36; 3-way-RM-ANOVA sex x rearing F(1,53) = 0.51, p = 0.47, η2 = 0.14). n = 13(F Ctrl), 16 (F ELA), 15 (M Ctrl), 16 (M ELA). These data have been replicated in three independent experiments. d Cue-potentiated startle was heightened in females (2-way-ANOVA sex F(1, 53) = 8.25, p = 0.005, η2 = 0.39). n = 13 (F Ctrl), 16 (F ELA), 15 (M Ctrl), 16 (M ELA). e Representative images of CeAL immunostained with c-Fos+ in CRF-IRES-Cre;Ai14tdT mice. f c-Fos + /DAPI was elevated in ELA mice following startle testing (2-way-ANOVA rearing F(1, 25) = 5.41, p = 0.02, η2 = 0.47) and was not influenced by sex (2-way-ANOVA sex F(1, 25) = 0.30, p = 0.58, η2 = 0.12) or an interaction (2-way-ANOVA sex x rearing F(1, 25) = 1.22, p = 0.27, η2 = 0.21). n = 7 (F Ctrl), 7 (F ELA), 8 (M Ctrl), 7 (F ELA). g c-Fos+CRF + /CRF+ was increased in ELA females (p = 0.003), but not males (p > 0.99; 2-way-ANOVA sex x rearing F(1, 25) = 5.75, p = 0.02, η2 = 0.47 with Bonferroni’s multiple comparisons). n = 7 (F Ctrl), 7 (F ELA), 8 (M Ctrl), 7 (F ELA). h c-Fos+CRF + /c-Fos+ was not impacted by sex, rearing or an interaction (2-way-ANOVA sex F(1,25) = 2.13, p = 0.15, η2 = 0.04; 2-way-ANOVA rearing F(1,25) = 1.23, p = 0.27, η2 = 0.07; 2-way-ANOVA sex x rearing F(1, 25) = 3.81, p = 0.06, η2 = 0.39). I Average startle was positively correlated with c-Fos+CRF + /c-Fos+ (Linear Regression: F(1, 25) = 14.83, R2 = 0.37, p = 0.0007). n = 7 (F Ctrl), 7 (F ELA), 8 (M Ctrl), 7 (F ELA). Plots depict individual mice and means ± SEM. *< 0.05, **< 0.01, ***< 0.001, ****< 0.0001. Source data are provided as a Source Data file. Figure 1a was created with BioRender.com and released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.

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