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Phospholipid scramblase 1 (PLSCR1) in villous trophoblast of the human placenta

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Abstract

A crucial factor for effective villous trophoblast fusion in the human placenta is the transient deregulation of plasma membrane phospholipid asymmetry leading to externalization of phosphatidylserine to the outer membrane leaflet. Screening of scramblase family members implicated in the collapse of phospholipid asymmetry revealed that phospholipid scramblase 1 (PLSCR1) is strongly expressed in villous trophoblast. Therefore, we assessed the putative role of PLSCR1 in villous trophoblast fusion. Spatio-temporal analysis in first trimester and term placenta showed abundant expression of PLSCR1 in syncytiotrophoblast, macrophages and endothelial cells, while it was virtually absent in villous cytotrophoblasts. For functional studies, BeWo cells, isolated primary term trophoblasts and first trimester villous explants were used. During forskolin-mediated BeWo cell differentiation, neither PLSCR1 mRNA nor protein levels showed significant changes. In contrast, when primary trophoblasts were stimulated with Br-cAMP, a decrease in PLSCR1 mRNA and protein expression was observed. To elucidate a role for PLSCR1 in syncytialization, we used RNA interference and a chemical scramblase inhibitor, R5421 (ethanedioic acid). Silencing of PLSCR1 using siRNA had no effects while inhibition of scramblase activity by R5421 increased GCM-1 mRNA expression, beta-hCG protein secretion and fusion rates of BeWo cells. In primary trophoblasts and villous explants, no effects of siRNA or R5421 treatment on fusion were detected. This study provides data on PLSCR1 localization and general expression in the human placenta. The data make it tempting to speculate on a role of PLSCR1 in negatively regulating trophoblast fusion.

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Acknowledgments

Primary trophoblast cells from isolated term placentas were obtained from the laboratory of Larry Guilbert, University of Alberta Edmonton, Canada, isolated by Bonny Lowen and Meghan R. Riddell. Experiments were performed in Denise G. Hemmings’ laboratory, University of Alberta. First trimester placenta samples were kindly provided by Andreas Glasner, Graz, Austria. Beta-hCG analyses with ADIVA Centaur XP Immunoassay System were performed at Labor Dr. Berghold—Medizinische Diagnostik, Graz, Austria. Measurements of beta-hCG with ELISA 1911 kit were generously carried out by Martina Mackova, University of Alberta. Analysis of platelet aggregation was performed at the Institute of Experimental and Clinical Pharmacology, Medical University of Graz with the kind help of Martina Ofner. This work was partly supported by the START funding program and the Franz-Lanyar-Foundation of the Medical University of Graz granted to Gerit Moser (Project # 358) and Martin Gauster (Project # 359). Martin Gauster was supported by the Austrian Science Fund (FWF): P23859-B19. Veronika M. Berghold and Julia Kremshofer were funded by the PhD program Molecular Medicine of the Medical University of Graz. Additionally, funding support was provided by the Canadian Institutes of Health Research (CIHR) to Denise G. Hemmings (MOP 123488).

Conflict of interest

The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.

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Authors and Affiliations

  1. Institute of Cell Biology, Histology and Embryology, Medical University of Graz, Harrachgasse 21/VII, 8010, Graz, Austria

    Veronika M. Berghold, Martin Gauster, Gerit Moser, Julia Kremshofer, Monika Siwetz, Monika Sundl & Berthold Huppertz

  2. Department of Obstetrics and Gynecology, University of Alberta, 227B Heritage Medical Research Centre, Edmonton, AB, T6G 2S2, Canada

    Veronika M. Berghold & Denise G. Hemmings

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  1. Veronika M. Berghold
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Correspondence to Berthold Huppertz.

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Berghold, V.M., Gauster, M., Hemmings, D.G. et al. Phospholipid scramblase 1 (PLSCR1) in villous trophoblast of the human placenta. Histochem Cell Biol 143, 381–396 (2015). https://doi.org/10.1007/s00418-014-1294-y

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