Molecular Human Reproduction, Vol. 6, No. 8, 743-749,
August 2000
© 2000 European Society of Human Reproduction and Embryology
Pregnancy |
Inhibin and activin production and subunit expression in human placental cells cultured in vitro
Department of Obstetrics and Gynecological Endocrinology, Université Catholique de Louvain, 1200 Brussels, Belgium
Abstract
Inhibins and activins are dimeric proteins, with each subunit being one of three related protein subunits (
, ßA or ßB). The mRNA levels of these subunits were studied quantitatively during in-vitro differentiation of human cytotrophoblast cells into syncytium, using Northern blot analysis and semi-quantitative reverse transcription–polymerase chain reaction (RT–PCR) analysis. The corresponding protein concentrations were determined by specific enzyme-linked immunosorbent assays for inhibin A, B, pro C and activin A in cellular protein extracts and culture medium (n = 5). Immunofluorescence studies showed syncytium formation after 48 h. The subunit was present before plating and increased at 48 h (P < 0.001) while the ßA subunit was weak before plating and increased at 24 h. The ßB subunit was not detected. With respect to corresponding protein synthesis, inhibin A ( + ßA) had risen after 48 h in cellular protein extract and after 72 h in culture medium, while activin A (ßA + ßB) was detected after 24 h, with no significant variations in culture medium. There was a good correlation between inhibin A and subunit expression (r = 0.736, P < 0.001), as well as between activin A and ßA subunit expression (r = 0.755, P < 0.001). This study showed that mRNA expression parallels protein synthesis of inhibin and activin in trophoblast cells. Inhibin A synthesis appears to be dependent on subunit mRNA expression, rather than on the ßA subunit which controls activin A synthesis. This study has also shown that isolated cytotrophoblast cells do not produce dimeric inhibin. However, during the transformation of cytotrophoblast cells into syncytium, ßA subunit mRNA expression may be an indicator of cell aggregation, while subunit mRNA expression may be an indicator of cell fusion.
activin/inhibin/mRNA/placenta/trophoblast
Notes
1 To whom correspondence should be addressed: Physiology of Human Reproduction Research Unit, Department of Obstetrics and Gynecological Endocrinology, Faculty of Medicine, OBST 5330, Avenue Em. Mounier 53, B-1200 Brussels, Belgium. E-mail: debieve{at}obst.ucl.ac.be
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