Mating-activated nitric oxide-producing neurons in specific brain regions in the female rat

Nitric oxide (NO)-containing neurons have been localized in various parts of the central nervous system including the hypothalamus. NO plays an important role in the regulation of reproductive activities including sexual behavior and pituitary hormone secretion. To test the hypothesis that NO-containing neurons in specific brain areas may respond to the stimulus of mating and participate in integrating the tactile information in the hypothalamus, this study used Fos as a marker of neuronal activity. Proestrous rats receiving intromissions (mated group) from males or mounts-without-intromission (mounted group) were sacrificed along with rats taken directly from their home cage (control group) 90 min after the beginning of mating or mounting. NOergic neurons were labeled by histochemical reaction for nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d). The presence of activated NO-producing (double-stained NADPH-d/Fos) neurons was quantitatively assessed in several brain areas before and after mating. The results showed that mating-with-intromissions induced a significant increase in the percentage of NADPH-d/Fos colabeled neurons in the medial preoptic area (mPOA) and the magnocellular component of the paraventricular nucleus (PVNm) compared to mounts-without-intromission or control treatment. Both mating and mounting induced Fos expression in NADPH-d-positive cells in the ventromedial nucleus of hypothalamus (VMN). In contrast, the expression of Fos in the NADPH-d-positive neurons in the supraoptic nucleus (SON) and the parvocellular portion of the paraventricular nucleus (PVNp) was not influenced by either mating or mounting although abundant NO-containing neurons were found in the two brain areas. The second experiment of the study examined whether NOergic neurons in these brain areas are influenced directly by estrogen by determining the number of NADPH-d-positive neurons that contained the estrogen receptor alpha (ERalpha), the classical ER. Double labeled NADPH-d/ERalpha neurons were observed in several brain areas including the mPOA and VMN while few, if any, NADPH-d-positive neurons in the SON, PVNm or PVNp contained ERalpha. The results suggest that the activated NOergic neurons in these brain areas may be involved in processing and integrating the mating stimulus. Further investigation is required to determine the physiological role of the mating-activated NOergic activity in specific mating-induced changes in reproductive neuroendocrinology.