Interrelationships between kisspeptin and FSH in control of porcine ovarian cell functions

Abstract

The existing knowledge of the direct action of kisspeptin on the ovary needs to be expanded. In our study, the direct effects of kisspeptin on basic ovarian cell functions and their response to FSH were examined. We studied the effect of kisspeptin alone (0, 1, 10, and 100 ng/mL) and of kisspeptin (1, 10, and 100 ng/mL) in combination with FSH (10 ng/mL) on cultured porcine granulosa cells. Markers of viability, proliferation (accumulation of proliferating cell nuclear antigen [PCNA] and cyclin B1), and apoptosis (accumulation of bax and caspase 3), as well as the release of steroid hormones and IGF-I were analyzed using the trypan blue exclusion test, quantitative immunocytochemistry, and ELISA. Addition of kisspeptin at lower doses (1 and 10 ng/mL) increased cell viability, the accumulation of PCNA and cyclin B1, decreased the accumulation of bax and caspase 3, and promoted release of progesterone, estradiol, and IGF-I, but not testosterone. A high dose (100 ng/mL) of kisspeptin had the opposite, inhibitory effect. The addition of FSH increased cell viability, proliferation, decreased apoptosis, and promoted progesterone, testosterone, estradiol, and IGF-I release. Kisspeptin at lower doses supported the stimulatory action of FSH on viability, PCNA and cyclin B1 accumulation, and release of progesterone and estradiol, promoted its inhibitory action on bax and caspase 3 accumulation, but did not modify its action on testosterone and IGF-I release. On the contrary, kisspeptin at a high dose inhibited and even reversed the FSH effect. FSH mimicked and promoted both the stimulatory and inhibitory action of kisspeptin on all examined ovarian functions besides IGF-I release. These observations show that kisspeptin can directly regulate basal ovarian cell functions. Furthermore, they demonstrate the functional interrelationships between kisspeptin and FSH in direct regulation of ovarian functions.