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Immunocytochemical localization of nuclear estrogen receptors and progestin receptors within the rat dorsal raphe nucleus

Estradiol and progesterone modulate central serotonergic activity; however, the mechanism(s) of action remain unclear. Recently, estradiol‐induced progestin receptors (PRs) have been localized within the majority of serotonin (5‐HT) neurons in the female macaque dorsal raphe nucleus (DRN; Bethea [1994] Neuroendocrinology 60:50–61). In the present study, we investigated whether estrogen receptors (ERs) and/or PRs exist within 5‐HT and/or non‐5‐HT cells in the female and male rat DRN and whether estradiol treatment alters the expression of these receptors. Young adult female and male Sprague‐Dawley rats were gonadectomized, and 1 week later, half of the animals received a subcutaneous Silastic implant of estradiol‐17β. Animals were transcardially perfused 2 days later with acrolein and paraformaldehyde, and sequential dual‐label immunocytochemistry was performed on adjacent sections by using either a PR antibody or an ERα antibody. This was followed by an antibody to either the 5‐HT‐synthesizing enzyme, tryptophan hydroxylase (TPH), or to the astrocytic marker, glial fibrillary acidic protein (GFAP). Cells containing immunoreactivity (ir) for nuclear ERs or PRs were identified within the rat DRN in a region‐specific distribution in both sexes. No colocalization of nuclear ER‐ir or PR‐ir with cytoplasmic TPH‐ir or GFAP‐ir was observed in either sex or treatment, indicating that the steroid target cells are neither 5‐HT neurons nor astrocytes. Females were found to have approximately 30% more PR‐labeled cells compared with males throughout the DRN (P < 0.05), but no sex difference was detected in the number of neurons demonstrating ER‐ir. In both sexes, 2 days of estradiol exposure decreased the number of cells with ER‐ir, whereas it greatly increased the number of cells containing PR‐ir in several DRN regions (P < 0.005). Collectively, these findings demonstrate the existence of nonserotonergic cells that contain nuclear ERs or PRs within the female and male rat DRN, including estradiol‐inducible PRs. These findings point to a species difference in ovarian steroid regulation of 5‐HT activity between the macaque and the rat, perhaps transsynaptically via local neurons in the rat brain. J. Comp. Neurol. 391:322–334, 1998. © 1998 Wiley‐Liss, Inc.