Benzodiazepine binding varies with stage of estrous cycle in unwashed membranes from mouse brain

The influence of the stage of the estrous cycle on binding of [³H]diazepam was examined in membranes from brains of female mice. In order to conserve endogenous factors such as progesterone, other steroids, or GABA, the assay was performed without the extensive washing procedures typically employed in measurements of benzodiazepine binding. Significant variations in the apparent maximal numbers of binding sites (B_max) were noted during the estrous cycle in both hypothalamus and cortex. The B_max measured in membranes from proestrus female mice was significantly higher than in membranes from mice at other stages in the estrous cycle. Variations in apparent equilibrium binding dissociation constants (K_d) were not statistically significant by stage of the estrous cycle. The demonstrated variations in binding suggest the existence of a factor which varies with the estrous cycle in female mice and modulates the activity of the GABA_A receptor complex.