Local production of the gonadotropic hormones in the rat ovary.

The gonadotropic hormones, luteinizing hormone (LH) and follicle-stimulating hormone (FSH) are synthesized by and released from the anterior pituitary in response to the hypothalamic gonadotropin-releasing hormone (GnRH) signaling. In the female, LH and FSH affect folliculogenesis, ovarian steroid production, oocyte maturation, ovulation and corpus luteum formation. We have recently studied the expression of GnRH and its receptor in the rat ovary and found organ-specific, estrous cycle-dependant, fluctuations. Subsequently, we wished to determine whether rat ovaries also express gonadotropic hormones. Using RT-PCR, we detected LHbeta, FSHbeta and the common alpha-subunit mRNA's in intact follicles, theca cells, corpora lutea and in meiotically competent and incompetent oocytes. Granulosa cells, however, express mRNA's for LHbeta and the common alpha-subunit, but not for FSHbeta. We cloned and sequenced the ovarian LHbeta transcript and found it to be longer (2.3kb) than the one produced by pituitary gonadotropes (0.8kb), due to a longer 5'-UTR. We studied the regulation of ovarian LHbeta mRNA in sexually immature female rats administered with pregnant mare serum gonadotropin (PMSG) and in adult cyclic rats. PMSG administration caused a significant decrease in LHbeta mRNA expression, detected by real-time PCR. Similarly, LHbeta mRNA levels were lower on estrous morning versus proestrous evening. Interestingly, ovarian content of LH remained unchanged following hypophysectomy, although ovarian weight was immensely reduced. Taken together, it seems probable that ovarian LH is heterologously/homologously regulated by pituitary, and possibly also by local gonadotropins. Thus, these findings may imply the existence of a local GnRH-gonadotropin axis in the mammalian ovary that may be involved in the management of processes that lead to ovulation.

Daily GnRH and GnRH-receptor mRNA expression in the ovariectomized and intact rat.

We recently described patterns of GnRH and GnRH receptor (GnRH-R) expression in the hypothalamus, pituitary and ovary throughout the rat estrus cycle. Here, we wished to distinguish between regulatory effects of ovarian factors and underlying circadian rhythmicity. We quantified GnRH and GnRH-R mRNA in the pituitary and hypothalamus of long-term ovariectomized (OVX) rats, at different times of day, using real-time PCR. Furthermore, we expanded our previous study of hypothalamic and pituitary GnRH and GnRH-R expression in intact rats by including more time points throughout the estrus cycle. We found different daily patterns of GnRH and GnRH-R expression in intact versus OVX rats, in both tissues. In the hypothalamus of OVX rats, GnRH mRNA peaked at 12, 16 and 20 h, whereas in the hypothalamus of intact rats we observed somewhat higher GnRH mRNA concentrations at 19 h on every day of the estrus cycle except proestrus, when the peak occurred at 17 h. In this tissue, GnRH-R fluctuated less significantly and peaked at 16 h in OVX rats. During the estrus cycle, we observed higher levels in the afternoon of each day except on estrus. In OVX rats, pituitary GnRH mRNA rose sharply at 9 h, with low levels thereafter. In these animals, pituitary GnRH-R also peaked at 9h followed by a second rise at 22 h. In intact rats pituitary GnRH was high at noon of diestrus-II and on estrus, whereas GnRH-R mRNA was highest in the evening of diestrus-II. This is the first demonstration of daily GnRH and GnRH-R mRNA expression patterns in castrated animals. The observed daily fluctuations hint at underlying tissue-specific circadian rhythms. Ovarian factors probably modulate these rhythms, yielding the observed estrus cycle patterns.

Differential gonadotropin-releasing hormone (GnRH) and GnRH receptor messenger ribonucleic acid expression patterns in different tissues of the female rat across the estrous cycle.

GnRH, the main regulator of reproduction, is produced in a variety of tissues outside of the hypothalamus, its main site of synthesis and release. We aimed to determine whether GnRH produced in the female rat pituitary and ovaries is involved in the processes leading to ovulation. We studied the expression patterns of GnRH and GnRH receptor (GnRH-R) in the same animals throughout the estrous cycle using real-time PCR. Hypothalamic levels of GnRH mRNA were highest at 1700 h on proestrus, preceding the preovulatory LH surge. No significant changes in the level of hypothalamic GnRH-R mRNA were detected, although fluctuations during the day of proestrus are evident. High pituitary GnRH mRNA was detected during the day of estrus, in the morning of diestrus 1, and at noon on proestrus. Pituitary GnRH-R displayed a similar pattern of expression, except on estrus, when its mRNA levels declined. Ovarian GnRH mRNA levels increased in the morning of diestrus 1 and early afternoon of proestrus. Here, too, GnRH-R displayed a somewhat similar pattern of expression to that of its ligand. To the best of our knowledge, this is the first demonstration of a GnRH expression pattern in the pituitary and ovary of any species. The different timings of the GnRH peaks in the three tissues imply differential tissue-specific regulation. We believe that the GnRH produced in the anterior pituitary and ovary could play a physiological role in the preparation of these organs for the midcycle gonadotropin surge and ovulation, respectively, possibly via local GnRH-gonadotropin axes.