Modulation of pp60c-src activity and cellular localization during differentiation of human trophoblast cells in culture.
The morphological and functional differentiation of human trophoblast cells ends with the formation of terminally differentiated multinucleated syncytial trophoblasts. This in vivo differentiation is mimicked in vitro during the primary culture of extravillous cytotrophoblasts: isolated mononuclear cytotrophoblasts aggregate and fuse to form syncytia. This in vitro differentiation is associated with an increase in epidermal growth factor receptor (EGF-R) expression and a transitory increase in E-cadherin expression during cell aggregation. In the present study, we investigated the expression of pp60c-src during morphological differentiation of trophoblast cells. Cultures were terminated at various time intervals and pp60c-src was analysed by immunocytochemistry using a specific antibody. In addition, pp60c-src was investigated by western blot analysis and its tyrosine kinase activity was measured concomitantly. In mononuclear cytotrophoblasts, pp60c-src was localized at cell-matrix contacts and during the aggregation of cytotrophoblasts, pp60c-src was distributed on the cell surface at points of cell-cell contact being colocalized with EGF-R and E-cadherin. The kinase activity of the pp60c-src protein increased significantly at day 2 when cells were completely aggregated and started to fuse, and remained elevated while cells underwent further differentiation. Inhibition of pp60c-src by herbimycin A at 0.25 to 1 microgram/ml during the first day of culture was associated with a decreased expression of tyrosine kinase activity of EGF-R and an increase in E-cadherin expression. These data suggest that pp60c-src is involved in the modulation of trophoblast cell aggregation and fusion leading to syncytial formation.
Rebut-Bonneton
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C
Laboratoire de Physiopathologie du Developpement, URA 1337 ENS, Paris, France.
Boutemy-Roulier
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Evain-Brion
D
D
D016227:Benzoquinones; D015820:Cadherins / Q000378:metabolism; D002449:Cell Aggregation; D002454:Cell Differentiation; D002459:Cell Fusion; D002478:Cells, Cultured; D066246:ErbB Receptors / Q000187:drug effects; D005260:Female; D015726:Giant Cells / Q000166:cytology / Q000378:metabolism; D006651:Histocytochemistry; D006801:Humans; D007365:Intercellular Junctions / Q000378:metabolism; D047029:Lactams, Macrocyclic; D011247:Pregnancy; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000187:drug effects / Q000378:metabolism*; D011809:Quinones / Q000494:pharmacology; D017828:Rifabutin / Q000031:analogs & derivatives; D014327:Trophoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
D016227:Benzoquinones; D015820:Cadherins; D047029:Lactams, Macrocyclic; D011809:Quinones; D017828:Rifabutin; C020754:herbimycin; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases; D016392:Proto-Oncogene Proteins pp60(c-src)