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First published online 2 July 2003
doi: 10.1242/jcs.00648
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Research Article |
1 Institut National de la Santé et de la Recherche Médicale, U427,
Paris, France
4 Laboratoire de Génétique Moléculaire, Faculté des
Sciences Pharmaceutiques et Biologiques, Université René
Descartes, Paris, France
2 Faculté des Sciences, Poitiers, France
3 Service d'Hormonologie, Hôpital Robert Debré, Paris, France
* Author for correspondence (e-mail: amalassi{at}pharmacie.univ-paris5.fr)
Accepted 2 May 2003
The syncytiotrophoblast is the principal component of the human placenta involved in feto-maternal exchanges and hormone secretion. The syncytiotrophoblast arises from the fusion of villous cytotrophoblasts. We recently showed that functional gap junctional intercellular communication (GJIC) is an important prerequisite for syncytiotrophoblast formation and that connexin 43 (Cx43) is present in cytotrophoblasts and in the syncytiotrophoblast. To determine whether Cx43 is directly involved in trophoblast fusion, we used an antisense strategy in primary cultures of human villous cytotrophoblasts that spontaneously differentiate into the syncytiotrophoblast by cell fusion. We assessed the morphological and functional differentiation of trophoblasts by desmoplakin immunostaining, by quantifying hCG (human chorionic gonadotropin) production and by measuring the expression of specific trophoblast genes (hCG and HERV-W). Furthermore, we used the gap-FRAP (fluorescence recovery after photobleaching) method to investigate functional GJIC. Cytotrophoblasts treated with Cx43 antisense aggregated and fused poorly. Furthermore, less HERV-W env mRNA, hCGß mRNA and hCG secretion were detected in Cx43 antisense-treated cytotrophoblasts than in cells treated with scrambled antisense. Treatment with Cx43 antisense dramatically reduced the percentage of coupled trophoblast cells. Taken together, these results suggest that Cx43 is directly involved in human trophoblast cell-cell communication, fusion and differentiation.
Key words: Cx43, Placenta, Herv-W, hCG, Cell-cell fusion
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