To study the regulatory relation of BDNF and FoxO1 in postpartum female mice, we conducted a correlation analysis between BDNF and FoxO1 levels in mPFC of CUS treated postpartum female mice, which indicated that total Bdnf mRNA levels were significantly positive correlated with FoxO1 in both mRNA (P = 0.010) and protein (P = 0.018) levels (Fig. 6a). Moreover, to determine whether FoxO1 can be regulated by BDNF in the mPFC, we injected AAV-Cre or AAV-GFP vectors bilaterally into the mPFC to delete Bdnf in the mPFC. Three weeks later, mPFC tissue was dissected for further analysis (Fig. 6b). Quantitative real-time PCR results demonstrated that total Bdnf mRNA levels were significantly reduced by AAV-mediated knockdown (Fig. 6c, P < 0.001). Furthermore, BDNF and FoxO1 protein levels were also significantly decreased in AAV-Cre-GFP-injected mice compared to AAV-GFP-injected control mice (Fig. 6d, BDNF: P < 0.001; FoxO1: P = 0.009). We also found a positive correlation between total Bdnf mRNA (P = 0.004) or protein (P = 0.018) levels with FoxO1 protein level in the AAV-Cre treated mice respectively (Fig. 6e).
