Molecular Human Reproduction, Vol. 8, No. 11, 1014-1022,
November 2002
© 2002 European Society of Human Reproduction and Embryology
Implantation and pregnancy |
Adhesiveness of human uterine epithelial RL95-2 cells to trophoblast: Rho protein regulation
1 Institut für Anatomie and 2 Institut für Pharmakologie, Universitätsklinikum Essen, D-45122 Essen, Germany
Embryo implantation involves adhesion of trophoblast cells to the epithelial lining of the endometrium. Using an in-vitro model to simulate this initial interaction, we previously reported that attachment of human trophoblast-like JAR spheroids to human uterine epithelial RL95-2 cells provokes a Ca2+ influx in RL95-2 cells depending on apically localized integrin receptors. Here, we demonstrate that adhesiveness of RL95-2 cells for JAR spheroids, measured by a centrifugal force-based adhesion assay, is dependent on Rho GTPases, most likely RhoA. Cellular expression and distribution of RhoA were studied by fluorescence confocal microscopy, focusing on the localization of RhoA and F-actin within the adhesion sites between JAR and RL95-2 cells. Contact areas contained high amounts of RhoA and F-actin fibres near the plasma membrane. To determine whether Rho GTPases may influence JAR cell binding, we treated RL95-2 cells with Clostridium difficile toxin A, which specifically inactivates Rho GTPases. Toxin A treatment changed the subcellular distribution of endogenous RhoA in RL95-2 cells and altered RhoA and F-actin colocalization. Adhesion of JAR spheroids to RL95-2 cells treated with toxin A was largely suppressed. These data indicate that Rho GTPases, most likely RhoA, play an important role in uterine epithelial RL95-2 cells for trophoblast binding, and suggest that RhoA may be involved in local signalling cascades during early embryo implantation in vivo.
actin cytoskeleton/adhesion/implantation/Rho GTPases/uterine epithelium
3 To whom correspondence should be addressed. E-mail: michael.thie{at}uni-essen.de
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