Biochemical characterization and modulation of LH/CG—receptor during human trophoblast differentiation

• DOI: 10.1002/jcp.20995
• Publication Date: 2007-04-26T05:47:28Z
• Authors: Guillaume Pidoux; Pascale Gerbaud; Vassilis Tsatsaris; Olivier Marpeau; Fatima Ferreira; Geri Meduri; Jean Guibourdenche; Josette Badet; Danièle Evain‐Brion; Jean‐Louis Frendo
• Keywords: 0301 basic medicine; cell fusion; placenta; hCG; syncytiotrophoblast; [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology; Cell Differentiation; cytotrophoblast; [SDV.BC]Life Sciences [q-bio]/Cellular Biology; Receptors, LH; Trophoblasts; 03 medical and health sciences; Gene Expression Regulation; Humans; RNA, Messenger; Cloning, Molecular; cell fusion.; [SDV.BDD]Life Sciences [q-bio]/Development Biology; hPL; Cells, Cultured

AbstractDue to the key role of the human chorionic gonadotropin hormone (hCG) in placental development, the aim of this study was to characterize the human trophoblastic luteinizing hormone/chorionic gonadotropin receptor (LH/CG‐R) and to investigate its expression using the in vitro model of human cytotrophoblast differentiation into syncytiotrophoblast. We confirmed by in situ immunochemistry and in cultured cells, that LH/CG‐R is expressed in both villous cytotrophoblasts and syncytiotrophoblasts. However, LH/CG‐R expression decreased during trophoblast fusion and differentiation, while the expression of hCG and hPL (specific markers of syncytiotrophoblast formation) increased. A decrease in LH/CG‐R mRNA during trophoblast differentiation was observed by means of semi‐quantitative RT‐PCR with two sets of primers. A corresponding decrease (∼60%) in LH/CG‐R protein content was shown by Western‐blot and immunoprecipitation experiments. The amount of the mature form of LH/CG‐R, detected as a 90‐kDa band specifically binding 125I‐hCG, was lower in syncytiotrophoblasts than in cytotrophoblasts. This was confirmed by Scatchard analysis of binding data on cultured cells. Maximum binding at the cell surface decreased from 3,511 to about 929 molecules/seeded cells with a kDa of 0.4–0.5 nM. Moreover, on stimulation by recombinant hCG, the syncytiotrophoblast produced less cyclic AMP than cytotrophoblasts, indicating that LH/CG‐R expression is regulated during human villous trophoblast differentiation. J. Cell. Physiol. 212: 26–35, 2007. © 2007 Wiley‐Liss, Inc.