Although GABA is the major inhibitory neurotransmitter in adult brain, it exerts depolarizing actions in developing neurons that include activation of voltage-gated calcium channels. The depolarizing actions of GABA serve an obvious trophic function, but the specific physiological significance of excitatory versus inhibitory GABA action has been largely ignored. We previously demonstrated that estradiol enhances the magnitude and duration of calcium influx through L-type voltage-gated calcium channels following GABA(A) receptor activation in neonatal hypothalamic neurons. This has led us to propose that GABA action represents a major divergence point in steroid-mediated sexual differentiation of rat brain. Presently, we examined sex differences in phosphorylation of the calcium-regulated transcription factor, cyclic AMP response element binding protein, following activation of the GABA(A) receptor with muscimol, in vivo. Muscimol given 30 min before killing significantly increased the number of neurons exhibiting phosphorylated cyclic AMP response element binding protein in newborn male hypothalamus and CA1 hippocampus but decreased phosphorylated cyclic AMP response element binding protein in most brain regions in females. Muscimol-induced increases in phosphorylated cyclic AMP response element binding protein in hypothalamus and hippocampus of newborn males were attenuated by pretreatment with the L-type voltage-gated calcium channel blocker, nimodipine, suggesting that calcium influx is involved in phosphorylation of cyclic AMP response element binding protein in neonate brain. Muscimol treatment had no effect on hypothalamic or hippocampal phosphorylated cyclic AMP response element binding protein levels in juvenile males and females. These results are consistent with a divergence in male and female rat brain in the calcium-mediated cellular response to muscimol that is restricted to the early neonatal period, a time critical for estradiol-mediated sexual differentiation.