title: Inhibition of Akt activity and calcium channel function coordinately drive cell-cell fusion in the BeWO choriocarcinoma placental cell line.
abstract: 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 (β-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.
authors.affiliation: Division of Reproduction, Warwick Medical School, University of Warwick, Coventry, United Kingdom.
authors.forename: Manu
authors.initials: M
authors.lastname: Vatish
authors.forename: Lydia
authors.initials: L
authors.lastname: Tesfa
authors.forename: Dimitris
authors.initials: D
authors.lastname: Grammatopoulos
authors.forename: Eijiro
authors.initials: E
authors.lastname: Yamada
authors.forename: Claire C
authors.initials: CC
authors.lastname: Bastie
authors.forename: Jeffrey E
authors.initials: JE
authors.lastname: Pessin
mesh_terms: D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000378:metabolism*; D002459:Cell Fusion; D045744:Cell Line, Tumor; D002822:Choriocarcinoma / Q000378:metabolism*; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000378:metabolism; D058534:Class I Phosphatidylinositol 3-Kinases / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D018613:Microscopy, Confocal; D010920:Placenta / Q000166:cytology* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*
chemical_list: D015220:Calcium Channels; D018997:Chorionic Gonadotropin, beta Subunit, Human; D005576:Colforsin; D000242:Cyclic AMP; D058534:Class I Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D002118:Calcium