Vatish, Manu, Tesfa, Lydia, Grammatopoulos, Dimitris K., Yamada, Eijiro, Bastie, Claire C. and Pessin, Jeffrey E.. (2012) Inhibition of Akt activity and calcium channel function coordinately drive cell-cell fusion in the BeWO choriocarcinoma placental cell line. PLoS ONE, Vol.7 (No.1). e29353. ISSN 1932-6203

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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.

Item Type:	Journal Article
Subjects:	Q Science > QP Physiology
Divisions:	Faculty of Medicine > Warwick Medical School > Metabolic and Vascular Health Faculty of Medicine > Warwick Medical School > Reproductive Health Faculty of Medicine > Warwick Medical School
Library of Congress Subject Headings (LCSH):	Cell hybridization, Choriocarcinoma, Placenta -- Physiology
Journal or Publication Title:	PLoS ONE
Publisher:	PLOS
ISSN:	1932-6203
Date:	19 January 2012
Volume:	Vol.7
Number:	No.1
Page Range:	e29353
Identification Number:	10.1371/journal.pone.0029353
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Funder:	National Institutes of Health (U.S.) (NIH), United States-United Kingdom Educational Commission, Winston Churchill Memorial Trust
Grant number:	DK082694 (NIH), DK020541 (NIH), DK081412 (NIH)
References:	1. Stadler B, Blattler TM, Franco-Obregon A (2010) Time-lapse imaging of in vitro myogenesis using atomic force microscopy. Journal of microscopy 237: 63–69. 2. Massarwa R, Carmon S, Shilo BZ, Schejter ED (2007) WIP/WASp-based actinpolymerization machinery is essential for myoblast fusion in Drosophila. Developmental cell 12: 557–569. 3. Malassine A, Cronier L (2002) Hormones and human trophoblast differentiation: a review. Endocrine 19: 3–11. 4. Knofler M, Vasicek R, Schreiber M (2001) Key regulatory transcription factors involved in placental trophoblast development–a review. Placenta 22 Suppl A: S83–92. 5. Chen EH, Olson EN (2005) Unveiling the mechanisms of cell-cell fusion. Science 308: 369–373. 6. Huppertz B, Bartz C, Kokozidou M (2006) Trophoblast fusion: fusogenic proteins, syncytins and ADAMs, and other prerequisites for syncytial fusion. Micron 37: 509–517. 7. Huppertz B, Borges M (2008) Placenta trophoblast fusion. Methods in molecular biology 475: 135–147. 8. Frendo JL, Olivier D, Cheynet V, Blond JL, Bouton O, et al. (2003) Direct involvement of HERV-W Env glycoprotein in human trophoblast cell fusion and differentiation. Molecular and cellular biology 23: 3566–3574. 9. Mori K, Nishimura M, Tsurudome M, Ito M, Nishio M, et al. (2004) The functional interaction between CD98 and CD147 in regulation of virus-induced cell fusion and osteoclast formation. Medical microbiology and immunology 193: 155–162. 10. Kudo Y, Boyd CA, Millo J, Sargent IL, Redman CW (2003) Manipulation of CD98 expression affects both trophoblast cell fusion and amino acid transport activity during syncytialization of human placental BeWo cells. The Journal of physiology 550: 3–9. 11. Ito N, Nomura S, Iwase A, Ito T, Kikkawa F, et al. (2004) ADAMs, a disintegrin and metalloproteinases, mediate shedding of oxytocinase. Biochemical and biophysical research communications 314: 1008–1013. 12. Lyden TW, Ng AK, Rote NS (1993) Modulation of phosphatidylserine epitope expression by BeWo cells during forskolin treatment. Placenta 14: 177–186. 13. Huppertz B, Frank HG, Kingdom JC, Reister F, Kaufmann P (1998) Villous cytotrophoblast regulation of the syncytial apoptotic cascade in the human placenta. Histochemistry and cell biology 110: 495–508. 14. Kudo Y, Boyd CA, Kimura H, Cook PR, Redman CW, et al. (2003) Quantifying the syncytialisation of human placental trophoblast BeWo cells grown in vitro. Biochimica et biophysica acta 1640: 25–31. 15. Borges M, Bose P, Frank HG, Kaufmann P, Potgens AJ (2003) A two-colour fluorescence assay for the measurement of syncytial fusion between trophoblastderived cell lines. Placenta 24: 959–964. 16. Wersto RP, Chrest FJ, Leary JF, Morris C, Stetler-Stevenson MA, et al. (2001) Doublet discrimination in DNA cell-cycle analysis. Cytometry 46: 296–306. 17. Keller PM, Person S, Snipes W (1977) A fluorescence enhancement assay of cell fusion. J Cell Sci 28: 167–177. 18. Hirsch E, Costa C, Ciraolo E (2007) Phosphoinositide 3-kinases as a common platform for multi-hormone signaling. The Journal of endocrinology 194: 243–256. 19. Kirby MS, Sagara Y, Gaa S, Inesi G, Lederer WJ, et al. (1992) Thapsigargin inhibits contraction and Ca2+ transient in cardiac cells by specific inhibition of the sarcoplasmic reticulum Ca2+ pump. J Biol Chem 267: 12545–12551. 20. Desforges M, Sibley CP (2010) Placental nutrient supply and fetal growth. The International journal of developmental biology 54: 377–390. 21. Crocker IP, Arthur P, Heazell AE, Baker PN (2007) The mitotic manipulation of cytotrophoblast differentiation in vitro. Placenta 28: 408–411. 22. Young BC, Levine RJ, Karumanchi SA (2010) Pathogenesis of preeclampsia. Annual review of pathology 5: 173–192. 23. Arnaudeau S, Holzer N, Konig S, Bader CR, Bernheim L (2006) Calcium sources used by post-natal human myoblasts during initial differentiation. Journal of cellular physiology 208: 435–445. 24. Ramos AJ, Cantero MR, Zhang P, Raychowdhury MK, Green A, et al. (2008) Morphological and electrical properties of human trophoblast choriocarcinoma, BeWo cells. Placenta 29: 492–502. 25. Fujita D, Tanabe A, Sekijima T, Soen H, Narahara K, et al. (2010) Role of extracellular signal-regulated kinase and AKT cascades in regulating hypoxiainduced angiogenic factors produced by a trophoblast-derived cell line. The Journal of endocrinology 206: 131–140.
URI:	http://wrap.warwick.ac.uk/id/eprint/42359

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