Fig. 3: Targeted Bmal1 knockout in the excitatory CaMK2a neurons of mPFC affects depression-like behavior and Homer1a expression. | Nature Communications

Fig. 3: Targeted Bmal1 knockout in the excitatory CaMK2a neurons of mPFC affects depression-like behavior and Homer1a expression.

From: Prefrontal cortex molecular clock modulates development of depression-like phenotype and rapid antidepressant response in mice

Fig. 3

a Experimental strategy used for region- and cell type-specific virally induced deletion of Bmal1. b Time course of the AAV microinjections into mPFC, baseline behavioral assessment in IntelliCage followed by CDM paradigm, ketamine treatment and test phase. c Relative mRNA expression of Bmal1 in mPFC at ZT06 (n = 10 mice per group, two-tailed Student’s t test: ***P < 0.001). d Representative images showing viral EGFP and EGFP-Cre expression in mPFC (scale bar, 500 µm) (left) and Bmal1 labeling (red) in EGFP and EGFP-Cre expressing cells (scale bar, 20 µm) (right). e Representative western blot and quantitative data of BMAL1 protein expression of in mPFC at ZT06 (n = 4 mice per group) and ZT18 (n = 3 EGFP, n = 5 Bmal1KO mice), two-tailed Student’s t test: *P < 0.05, **P < 0.01). f Immobility time during the induction and test phase FST of Bmal1-floxed mice mPFC bilaterally injected with control CaMK2a-EGFP or CaMK2a-Cre AAVs. (n = 10 mice per group, repeated measures two-way ANOVA with Bonferroni post hoc test: *P < 0.05 and #P < 0.05, ##P < 0.01, ###P < 0.001 vs day 1). g Immobility time during baseline and test phase TST (n = 10 mice per group, repeated measures two-way ANOVA with Bonferroni post hoc test: *P < 0.05). h Sucrose preference assessed in IntelliCage during baseline and test phase (n = 5 mice, repeated measures two-way ANOVA with Bonferroni post hoc test: **P < 0.01 and #P < 0.05, ##P < 0.01, ###P < 0.001 vs baseline). i Relative mRNA expression of clock genes and Homer1a in mPFC at ZT06 of control (n = 4), CDM (n = 5) and CDM 24 h post ketamine mice (n = 5) (two-way ANOVA with Bonferroni post hoc test: *P < 0.05, **P < 0.01, ***P < 0.001). j Immobility time of FST and TST during the test phase of CaMK2a-EGFP/Bmal1KO mice 24 h post single 3 mg/kg ketamine i.p. injection (KET) (n = 10, two-way ANOVA with Tukey’s post hoc test: *P < 0.05). k Bmal1KO in mPFC blocks Homer1a modulation by stress and ketamine, and thus inhibits the development of depression-like behavior and the antidepressant response. Data are presented as mean ± SEM and the individual data points are depicted. See also Supplementary Fig. 3 and Supplementary Data 1. Source data are provided as a Source Data file.

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