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. Author manuscript; available in PMC: 2020 Jan 9.
Published in final edited form as: Nat Neurosci. 2018 May 21;21(6):803–807. doi: 10.1038/s41593-018-0151-z

Fig. 3 |. CRH in CAMK2A neurons regulates anxiety, fear memory expression and dopamine release in the prefrontal cortex.

Fig. 3 |

a, Schematic illustration of the targeted Crh allele (Crhflox); details in Supplementary Fig. 7; UTR, untranslated region. b, c, In situ hybridization showing Crh mRNA deletion pattern in CrhCKO-Camk2a mice; quantifications from four independent experiments in Supplementary Fig. 8b. d, Dark-light box test: lit zone time (t(23) = 2.6, *P = 0.018) and lit entries (t(23) = 1.9, P = 0.068); unpaired two-tailed t-test, n = 12 CrhCtrl, 13 CrhCKO-Camk2a. e, Open arm time (%) in the EPM (t(18) = 2.4, P= 0.031; unpaired two-tailed t-test, n = 11 CrhCtrl, 9 CrhCKO-Camk2a). f, Marble burying test (t(31) = 2.8, **P = 0.009; unpaired two-tailed t-test, n = 16 CrhCtrl, 17 CrhCKO-Camk2a). g, Cued fear conditioning (repeated-measures ANOVA time × group interaction: F(6156) = 3.88, P = 0.0012; Bonferroni post hoc test, ⋆ P<0.05, ***P< 0.0001; n = 13 CrhCtrl, 15 CrhCKO<amk2a). h, In vivo microdialysis showing decreased dopamine release in the prefrontal cortex of CrhCKO-Camk2a mice under baseline conditions and following footshock (FS) stress (repeated-measures ANOVA genotype effect: F(117) = 7.14, ⋆ P = 0.02; n = 10 CrhCtrl, 9 CrhCKO-Camk2a). i, Cre-dependent expression of a constitutively active CRHR1-EGFP fusion construct (AAV-Ef1a::DIO-(CA)CRHR1-EGFP) in VTA neurons of Crhr1-ires-Cre mice (representative images from three independent experiments; details in Supplementary Figs. 11 and 12). Littermate controls were injected with AAV-Ef1a::DIO-mCherry. j-m, Crhr1-ires-Cre mice expressing (CA)CRHR1 in the VTA exhibit decreased anxiety in the dark-light box test (j; lit zone time: U = 14, *P= 0.043; lit zone entries: U = 20, P= 0.1; Mann-Whitney U test two-tailed; n = 7 Ctrl, 10 (CA)CRHR1), EPM (k; t(15) = 2.2, *P= 0.043; unpaired two-tailed t-test; n = 7 Ctrl, 10 (CA)CRHR1) and marble burying test (l; t(17) = 4.3, ***P = 0.005; unpaired two-tailed t-test; n = 8 Ctrl, 11 (CA) CRHR1) without showing alterations in cued fear conditioning (m; repeated-measures ANOVA time × group interaction: F(6102) = 0.45, P = 0.84; n = 8 Ctrl, 11 (CA)CRHR1). Anterior dorsal BNST, adBNST; anterior ventral BNST, avBNST; tyrosine hydroxylase, TH. Error bars represent s.e.m.