Effect of orexin A on the release of GnRH-stimulated gonadotrophins from cultured pituitary cells of immature and mature female rats.

Orexin A (OxA), also known as hypocretin 1, is a regulatory neuropeptide involved in the control of various autonomic and neuroendocrine functions. It appears to have a significant impact on the regulation of trophic hormones secretion by influencing the hypothalamus and the pituitary. Orexin A acts through two types of receptor found in the pituitary. This suggests the possibility of direct action of OxA at the adenohypophysis level. The aim of this study was to investigate the direct effect of OxA on GnRH (gonadotrophin-releasing hormone)-stimulated LH and FSH secretion from cultured pituitary cells of sexually immature and mature female rats. Anterior pituitary cells obtained from immature and mature female rats (ovariectomized, and ovariectomized and treated with estradiol) were incubated with 10(-10)M or 10(-7)M orexin A for 1 hour and 4h and the effect on GnRH-stimulated (10(-9)M or 10(-6)M) LH and FSH release was examined. The concentrations of secreted gonadotrophins in the culture media were determined by RIA methods. Orexin A significantly inhibited GnRH-stimulated FSH release from pituitary cells isolated from immature female rats, whereas in cells of mature ovariectomized animals, the effect of OxA was dependent on the stimulatory dose of GnRH. When the cells were stimulated with a low dose of GnRH, orexin A inhibited the secretion of gonadotrophins, but when a high dose of GnRH was used, orexin A increased mainly the release of LH. In cultured pituitary cells from ovariectomized, estrogenized mature rats, orexin A inhibited the secretion of LH if the cells were stimulated with a high dose of GnRH. In conclusion, the results of this study revealed that orexin A may modify the sensitivity of gonadotrophic cells to GnRH, and its effect depends on the maturity and estrogen status of the rats from which the cells are isolated.