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Adenosine 3',5'-Monophosphate, Prostaglandins, and Epinephrine Stimulate the Secretion of Immunoreactive Gonadotropin-Releasing Hormone from Cultured Human Placental Cells^*

FELICE PETRAGLIA, ALAN T. W. LIM and WYLIE VALE

Clayton Foundation Laboratories for Peptide Biology, The Salk Institute for Biological Studies LaJolla, California 92037

Address requests for reprints to: Dr. Felice Petraglia, Peptide Biology Laboratory, the Salk Institute for Biological Studies, P.O. Box 85800, San Diego, California 92138.

Cell culture and biochemical procedures were used to identify and study the possible mechanisms regulating the secretion of GnRH-like immunoreactivity (GnRH-LI) from human placenta. Monolayer primary cultures of trophoblasts were established after mechanical and enzymatic dispersion of normal human term placenta. The cultured cells stained immunocytochemically positive with anti-GnRH serum, and GnRH-LI extracted from the cells eluted from high performance liquid chromatography with the same retention time as authentic GnRH. One week after plating, exposure to high concentrations of K⁺ or to various doses of veratridine, a Na⁺ ionophore, increased GnRH-LI release into the culture medium. This effect was reversed by Ca²⁺ antagonists (cobalt, EGTA, and verapamil). Dibutyrylcyclic AMP, forskolin, theophylline, and theobromine also increased GnRH-LI concentrations in the medium of cultured placental cells in a dose-related manner, as did prostaglandins E₂ and F₂ and epinephrine. The effect of epinephrine on GnRH-LI concentrations was mimicked by isoproterenol and reversed by propranolol, suggesting an effect mediated by β-adrenergic receptors. These results indicate that GnRH-LI release from cultured human placental cells is stimulated by the opening of ionic channels and activation of the adenylate cyclase/cAMP system, and that prostaglandins and epinephrine may be involved in the regulation of GnRH-LI release from human placenta.

^* This work was supported by USPHS Grant HD-13527 and was conducted in part by the Clayton Foundation for Research, California Division.

Supported in part by the Consiglio Nazionale delle Ricerche (pos. 350.542). Permanent address: Department of Obstetrics and Gynecology, University of Modena School of Medicine, Via del Pozzo 71, 41100 Modena, Italy.

C. J. Martin fellow supported by the National Health and Medical Research Council of Australia.

Senior Clayton Investigator.

Received November 10, 1986.

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