The actions of androgens in both peripheral and central tissues are linked in part to their ability to specifically bind and activate androgen receptors (ARs). ARs have been well studied in the rat hypothalamus and peripheral reproductive tissues, where they are directly involved in endocrine feedback mechanisms and reproduction. Previous studies revealed relatively high levels of AR and AR messenger RNA (mRNA) in the rat hippocampus; however, the action of androgen in this brain region remains unclear. To begin to address this issue, we used a multidisciplinary approach to quantitate hippocampal AR and AR mRNA levels and investigate their regulation after various hormonal manipulations. In vitro binding assays revealed a single, saturable, high affinity binding site for androgen in hippocampal cytosols. The expression of AR mRNA in the intact adult male rat hypothalamus and hippocampus was demonstrated using reverse transcription-polymerase chain reaction and quantified using a ribonuclease protection assay. Comparable levels of AR mRNA were found in the hippocampus and hypothalamus. In addition, in situ hybridization analysis revealed a unique distribution of AR mRNA in the hippocampus. AR mRNA was found predominately in the CA1 pyramidal cells, which form the major signal output of the hippocampal trisynaptic circuit. Reverse transcription-polymerase chain reaction of total RNA from microdissected hippocampal regions confirmed this distribution. Ribonuclease protection assay demonstrated a significant decrease in the AR mRNA content of the hippocampus in animals killed 4 days after castration or in intact rats after four daily injections of the AR antagonist, flutamide (15 mg/animal), compared to that in intact controls (P < 0.01). In contrast, a 35% increase (P < 0.05) in the hippocampal AR mRNA content was found in old (22-month-old) compared to young (5-month-old) male rats. In both cases, [3H]dihydrotestosterone binding to the cytosolic preparation did not parallel the changes observed in the AR mRNA content. Taken together, these data demonstrate that hippocampal cells containing AR can respond to circulating androgen to alter AR gene expression. Furthermore, AR mRNA autoregulation appears to be both age and tissue specific and does not directly follow the regulatory patterns described for other steroid hormone receptors found in the hippocampus.