Involvement of phospholipase C and intracellular calcium signaling in the gonadotropin-releasing hormone regulation of prolactin release from lactotrophs of tilapia ( Oreochromis mossambicus )

Gonadotropin-releasing hormone (GnRH) is a potent stimulator of prolactin (PRL) secretion in various vertebrates including the tilapia, Oreochromis mossambicus. The mechanism by which GnRH regulates lactotroph cell function is poorly understood. Using the advantageous characteristics of the teleost pituitary gland from which a nearly pure population of PRL cells can be isolated, we examined whether GnRH might stimulate PRL release through an increase in phospholipase C (PLC), inositol triphosphate (IP 3 ), and intracellular calcium (Cai2+) signaling. Using Cai2+ imaging and the calcium-sensitive dye fura-2, we found that chicken GnRH-II (cGnRH-II) induced a rapid dose-dependent increase in Cai2+ in dispersed tilapia lactotrophs. The Cai2+ signal was abolished by U-73122, an inhibitor of PLC-dependent phosphoinositide hydrolysis. Correspondingly, cGnRH-II-induced tPRL 188 secretion was inhibited by U-73122, suggesting that activation of PLC mediates cGnRH-II’s stimulatory effect on PRL secretion. Pretreatment with 8-( N, N -diethylamino)octyl-3,4,5-trimethoxybenzoate hydrochloride (TMB-8), an inhibitor of Ca 2+ release from intracellular stores, impeded the effect of cGnRH-II on Cai2+. To further address the possible involvement of intracellular Ca 2+ stores, IP 3 concentrations in the tilapia rostral pars distalis (RPD containing 95–99% PRL cells) was determined by a radioreceptor assay. We found that GnRH-II induces a rapid (<5 min) and sustained increase in IP 3 concentration in the RPD. Secretion of tPRL 188 in response to cGnRH-II was suppressed by Ca 2+ antagonists (TMB-8 and nifedipine). These data, along with our previous findings that show PRL release increases with a rise in Cai2+, suggest that GnRH may elicit its PRL releasing effect by increasing Cai2+. Furthermore, the rise in Cai2+ may be derived from PLC/IP 3 -induced mobilization of Ca 2+ from intracellular stores along with influx through L-type voltage-gated Ca 2+ channels.