Para/autocrine regulation of estrogen receptors in hippocampal neurons

Previous studies have shown that estrogens, originating from ovaries, have a wide variety of estrogen receptor (ER)-mediated effects in the hippocampus. In the present study, we have investigated whether estrogens, which are synthesized in the hippocampus, could induce these effects as well. As a parameter, we used ER expression in response to estrogen synthesis, because estrogen receptors are ligand-inducible transcription factors. The experiments were carried out with cultures of isolated adult rat hippocampal cells, which contained about 95% neurons and about 5% oligodendrocytes in serum-free and steroid-free medium. Hippocampal neurons express both estrogen receptor isoforms (ERalpha and ERbeta), as shown by reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization. The release of estrogens by hippocampal neurons was quantified by radioimmunoassay (RIA). The ER isoforms (alpha and beta) were studied by semiquantitative immunocytochemical image analysis. Hippocampal cells precultured for 4 days were found to synthesize 17beta-estradiol for the next 8 days. This synthesis was completely inhibited by letrozol, an aromatase inhibitor. Inhibition of estrogen synthesis by letrozol induced a significant decrease in ERalpha expression, but an increase in ERbeta. As a control, supplementation of the medium with 17beta-estradiol resulted in a significant increase of ERalpha expression, whereas ERbeta was downregulated. Our findings provide evidence for a de novo synthesis of estrogens in the hippocampus, differential regulation of estrogen receptor isoforms by estrogen and consequently for a para/autocrine loop of estrogen action in the hippocampus.