Maternal care variations associate with DNA methylation of the glucocorticoid receptor gene, Nr3c1, in hippocampus at a nerve-growth factor-inducible protein 1 binding site. Epigenetic regulation of brain-derived neurotrophin factor is affected by early stress. These systems contribute to anxiety and fear. Early stress has sex-dependent effects perhaps reflecting sex differences in maternal care. Altering litter gender composition affects maternal behavior and DNA methylation levels of another gene in hippocampus and nucleus accumbens (NAc). We now test if DNA methylation levels of Nr3c1, Egr1, and Bdnf differ by litter composition or sex. Rats from mixed- or single-sex litters were tested for anxiety and fear on postnatal day 35. Brain tissues were collected and analyzed using direct sequencing methods. Females showed hypermethylation of Nr3c1 of hippocampal DNA and litter composition modified sex effects on methylation of Egr1 in NAc. Few differences were seen for Bdnf. LGC modified some sex differences in behavior.
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