%0 Journal Article
%T Inhibition of Akt Activity and Calcium Channel Function Coordinately Drive Cell-Cell Fusion in the BeWO Choriocarcinoma Placental Cell Line
%A Manu Vatish
%A Lydia Tesfa
%A Dimitris Grammatopoulos
%A Eijiro Yamada
%A Claire C. Bastie
%A Jeffrey E. Pessin
%J PLOS ONE
%D 2012
%I Public Library of Science (PLoS)
%R 10.1371/journal.pone.0029353
%X To establish a simple and quantitative live cell fusion assay for placental syncytialization, we generated stable GFP and dsRed expressing fusogenic BeWo cell lines. Fluorescent Activated Cell Sorting was shown to provide a quantitative determination of forskolin (cAMP-mediated) fusion in a time and concentration dependent manner consistent with the increased secretion of beta human chorionic gonadotrophin (¦Â-HCG) and appearance of multi-nucleated cells. Analyses of the fusion process demonstrated that in addition to increased cAMP levels, simultaneous reduction of intracellular calcium and inhibition of Type 1 phosphatidylinositol 3 kinase (PI3K)/Akt signaling also resulted in cell fusion. Although individual blockade of calcium channel function or PI3K/Akt signaling was without effect, the combination with forskolin resulted in a potentiation of cell fusion. These data demonstrate syncytialization is a complex process that depends upon the regulation of distinct signaling inputs that function in concert with each other.
%U http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0029353