Identification of genes and pathways associated with chronic restraint stress-induced anxiodepressive-like behaviors in female mice by transcriptomic and genome-wide DNA methylation combinational-analysis.
Women are at a higher risk for depression and anxiety than men, yet research into the biological mechanisms and pharmacological treatments for female-specific mental disorders, such as perinatal and perimenopausal depression, remains insufficient. DNA methylation, a key epigenetic modification, has been implicated in the pathogenesis of depression. In this study, we exposed female mice to chronic restraint stress (CRS) for 2 weeks, which induced both depression-like and anxiety-like behaviors. We then collected genomic DNA and total RNA from the medial prefrontal cortex (mPFC), a brain region critically involved in depression pathophysiology, and performed RNA sequencing (RNA-Seq) and whole-genome enzymatic methylation sequencing (EM-seq) to investigate stress-related transcriptional and epigenetic changes. Transcriptomic analysis identified 105 upregulated and 71 downregulated genes in the mPFC of CRS-exposed mice. Notably, CRS increased the expression of genes associated with the complement and coagulation cascades, while downregulating the expression of genes encoding neurotransmitter receptors, their downstream signaling regulators, and markers of neuronal activity. Epigenetic profiling revealed 6,848 hypermethylated and 4,530 hypomethylated regions, with CRS altering both CG- and CHH-type DNA methylation. Integrative analysis uncovered 13 downregulated genes with significant hypermethylation, most of which were implicated in neuronal activity regulation, and 8 upregulated genes with hypomethylation. Our findings suggest that CRS-induced DNA hypermethylation at CG and CHH sites in the female mPFC may contribute to stress-induced transcriptional dysregulation, potentially driving anxiodepressive-like behaviors. These results provide new insights into the epigenetic mechanisms underlying sex-specific stress responses.