The “play transcriptome” is sex-specific in the juvenile rat medial amygdala: a role for inhibitory neurons

Social play is a well-conserved, dynamic behavior known to be sexually differentiated. In most species, males play more than females, a sex difference largely driven by the medial amygdala (MeA) yet the full mechanism establishing this bias is unknown. Here, we explore “the transcriptome of playfulness” in both sexes for the first time, demonstrating that the transcriptomic profile in the juvenile rat MeA associated with playfulness is markedly distinct in males and females. Parallel single-cell RNA-sequencing experiments from newborn rats suggest that inhibitory neurons drive this sex difference. Furthermore, we show that inhibitory neurons comprise the majority of play-active cells in the juvenile MeA, with males having more play-active cells than females, of which a greater proportion are GABAergic. Through integrative bioinformatic analyses, we further explore the expression, function, and cell-type specificity of key play-associated gene modules, providing valuable insight into the sex-biased mechanisms underlying this fundamental social behavior. In this study, we generated single-cell RNA-seq (scRNA-seq) data from the amygdala of 4-day-old male and female rats. Amygdala tissue from 3 animals per sex were pooled into each of two samples (Male_A, Male_B, Female_A, Female_B) submitted for sequencing.