During development, estrogen has a variety of effects on morphological, biochemical and electrophysiological properties of hippocampal neurons. Correspondingly, estrogen receptor (ER) binding and mRNA increase transiently in the developing hippocampus. In this study, we used immunocytochemistry to determine the localization of the ER alpha subtype in the developing rat hippocampus. Nuclear staining was present in pyramidal cells and some interneurons of the CAI and CA3 regions of the developing rat hippocampus. Little or no immunoreactivity was observed in postnatal day (P)0 animals (day of birth = P0), however, be.-inning on P4, ER alpha -immunoreactivity (ER alpha -ir) was visible and reached maximal levels by P10. These levels subsequently declined to low levels so that by P15, levels approximated those of adult females. Western blot analysis confirmed that this antibody recognized a 67 kDa protein, characteristic of the full-length ER alpha. protein, in the hippocampus and pituitary. Furthermore, most of the ER alpha -immunopositive cells in the hippocampus were located in the pyramidal cell layer, and did not co-localize appreciably with gamma -aminobutyric acid (GABA) at any age examined. We conclude, based on the immunocytochemical localization of ER alpha, thatt the effects of estrogen on biochemistry and morphology of the developing hippocampus may be direct through the ER alpha subtype in hippocampal pyramidal cells. (C) 2001 Elsevier Science BY All rights reserved.