Molecular Human Reproduction, Vol. 6, No. 12, 1119-1130,
December 2000
© 2000 European Society of Human Reproduction and Embryology
Pregnancy |
Calcium influx in human uterine epithelial RL95-2 cells triggers adhesiveness for trophoblast-like cells. Model studies on signalling events during embryo implantation
1 Max-Planck Institut für molekulare Physiologie, D-44227 Dortmund, and 2 Institut für Anatomie, Universitätsklinikum, D-45122 Essen, Germany
Abstract
RL95-2 is a human uterine epithelial cell line that exhibits adhesion competence on its apical surface for trophoblast-like JAR cells. Using confocal microscopy and an adhesion assay we have found that changes in intracellular free calcium ([Ca2+]i) in RL95-2 cells are involved in binding of JAR spheroids. Impact of spheroids upon, and movement of spheroids across, monolayers of RL95-2 cells produced a transient increase in [Ca2+]i. Pretreatment of RL95-2 cells with the Ca2+ channel inhibitor, diltiazem, reduced the [Ca2+]i increase. Interestingly, resting of JAR spheroids on RL95-2 cells caused no detectable alterations in [Ca2+]i although cell–cell bonds were formed during prolonged contact. However, separation of established bonds did produce an increase in [Ca2+]i which could be reduced by the Ca2+ channel blocker, SKF-96365, but not by diltiazem. SKF-96365 also reduced adhesion of JAR spheroids to RL95-2 cells. In all experiments, the increase in [Ca2+]i was due to influx from the external medium, as it could be blocked both by removing extracellular Ca2+ and by nickel. These results suggest that the plasma membrane of uterine RL95-2 cells contains two types of Ca2+ channels that are involved in trophoblast adhesion, i.e. diltiazem-sensitive channels contributing to initiation of JAR cell binding and SKF-96365-sensitive channels participating in a feedback loop that controls the balance of bonds.
calcium influx/cell adhesion/implantation/polarized phenotype/uterine epithelium
Notes
3 To whom correspondence should be addressed. E-mail: michael.thie{at}uni-essen.de
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