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Vasoactive intestinal peptide increases intracellular cyclic amp and gonadotropin alpha gene activity in jeg 3 syncytial trophoblasts constraints posed by desensitization

, : Vasoactive intestinal peptide increases intracellular cyclic amp and gonadotropin alpha gene activity in jeg 3 syncytial trophoblasts constraints posed by desensitization. Journal of Biological Chemistry 265(18): 10274-10281

Expression of the human chorionic gonadotropin (hCG)-.alpha. gene in placental trophoblasts is markedly stimulated by cAMP, a property preserved in a reporter plasmid containing its cAMP response elements (CREs) linked to the chloramphenicol acetyltransferase coding sequence (CRE.alpha.CAT). In search of a potential physiologic regulator of hCG gene expression via cAMP, we found that JEG-3 syncytial trophoblast cells have specific binding sites for vasoactive intestinal peptide (VIP) with dissociation constant of 1 nM. VIP maximally increased the transient expression of CRE.alpha.CAT and the expression of endogenous hCG-.alpha. mRNA in JEG-3 cells by 4- and 9-fold, respectively. Exposure of JEG-3 cells to 30 min, but cAMP rapidly declined thereafter. As a consequence of this desensitization, the effect of VIP on stimulation of both CRE.alpha.CAT and endogenous hCG-.alpha. and hCG-.beta. mRNA levels more closely resembled that of forskolin or 8-br-cAMP at time points much less than 24 h. Moreover, transient exposure to 8-br-cAMP was much less effective than 24 h of continuous incubation on CRE.alpha.CAT activity. We conclude that VIP rapidly increases cAMP content and activates hCG-.alpha. gene expression in JEG-3 cells, but sustained elevations in cAMP are necessary for maximal accumulation of this CRE-regulated gene product.


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