Fig. 3: CeAL CRF+ neurons were necessary for within-session startle but not freezing, with a lasting impact on both startle and freezing. | Nature Communications

Fig. 3: CeAL CRF+ neurons were necessary for within-session startle but not freezing, with a lasting impact on both startle and freezing.

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

Fig. 3

a Experimental design. b CeAL CRF + neuron chemogenetic inhibition reduced startle independent of noise alone (NA) or CS + trials (2-way-RM-ANOVA virus F(1, 43 = 29.42, p = 2.4e-6, η2 = 1.45). n = 23 (mCh), 22 (hM4di). c Average startle (Blocks 1 & 2) was reduced independent of rearing or sex (3-way-ANOVA virus F(1, 37) = 27.96, p = 5.7e-6, η2 = 0.86; rearing F(1, 37) = 0.75, p = 0.39, η2 = 0.14; sex F(1, 37) = 0.90, p = 0.34, η2 = 0.15). 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). d CeAL CRF + inhibition did not impact percent freezing (2-way-ANOVA virus F(1, 43) = 0.01 p = 0.89, η2 = 0.03). n = 23 (mCh), 22 (hM4di). e Average freezing was not impacted by CRF + neuron inhibition. Freezing was greater in females and lower in ELA reared mice (3-way-ANOVA virus F(1, 37) = 0.00009, p = 0.97, η2 = 0.02; sex F(1, 37) = 4.99, p = 0.03, η2 = 0.36; rearing F(1, 37) = 3.86, p = 0.05, η2 = 0.31). f CRF + neuron inhibition did not impact distance traveled in the open field (Unpaired, Two-Tailed: t = 0.37, df = 43, p = 0.71, d = 0.11). 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). g CRF + neuron inhibition did not impact the frequency of center entries (Unpaired, Two-Tailed: t = 0.19, df = 43, p = 0.84, d = 0.05). n = 23 (mCh), 22 (hM4di). h CRF + inhibition did not impact the time spent in the center (Unpaired, Two-Tailed: t = 0.75, df = 43, p = 0.45, d = 0.10). n = 23 (mCh), 22 (hM4di). i Prior CRF + neuron inhibition impaired startle during re-exposure (2-way-RM-ANOVA virus F(1, 43) = 21.50, p = 3.2e-5, η2 = 0.96). n = 23 (mCh), 22 (hM4di). j Prior CRF + neuron inhibition increased freezing during habituation (p = 0.0002) and Block 1 (p = 0.003) of re-exposure, but not during CS + (p = 0.65; (2-way-RM-ANOVA virus F(1, 43) = 59.06, p = 0.01, η2 = 0.70; cue x virus F(3, 129) = 26.44, p = 2.1e-13, η2 = 0.78). n = 23 (mCh), 22 (hM4di). All plots depict individual mice and means ± SEM. *< 0.05, **< 0.01, ***< 0.001. Source data are provided as a Source Data file. Figure 3a created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.

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