Effects of prostaglandin E2, forskolin and cholera toxin on cAMP production and in vitro LH-RH release from the rat hypothalamus.

The present study attempts to elucidate the possible role of adenosine 3',5'-monophosphate (cAMP) and prostaglandin E2 (PGE2) in the function of the neural luteinizing hormone-releasing hormone (LH-RH) apparatus. To this end, in vitro LH-RH release from superfused hypothalamic fragments and cAMP production by hypothalamic P2 membrane fractions were measured. Immature female rats (day 28) were ovariectomized and implanted with Silastic capsules containing estradiol (235 micrograms/ml). Two days later, animals were sacrificed and the mediobasal hypothalamic preoptic area (hypothalamic units or fragments) were removed. To examine in vitro LH-RH release from superfused hypothalamic fragments, effluents were collected into tubes on ice at 10-min intervals and LH-RH concentration was determined by radioimmunoassay (RIA). Following a 50-min control period, a step-wise increment in several doses of PGE2 (each dose for a 50-min interval) evoked a dose-related increase in LH-RH release. PGE2 induced significant (P less than 0.01) increments in LH-RH release at doses of 5.68 X 10(-7), 5.68 X 10(-6), and 5.68 X 10(-5) M, respectively. When adenylate cyclase activators, such as forskolin and cholera toxin were infused in a step-wise manner (each dose for a 50-min interval) following a 50-min control period, a dose-related increase in LH-RH release was also obtained; forskolin and cholera toxin significantly (P less than 0.01) stimulated LH-RH release at doses of 1 X 10(-4) and 5.4 X 10(-10) M, respectively. These two substances were ineffective in stimulating LH-RH release when hypothalamic fragments were superfused in calcium-free plus EGTA (10 mM) containing medium.(ABSTRACT TRUNCATED AT 250 WORDS)