Proceedings of The Physiological Society

University of Manchester (2010) Proc Physiol Soc 19, PC246

Poster Communications

Sphingosine-1-phosphate receptor expression in extravillous trophoblasts

K. Alsaghir1, D. Adlam1, M. Westwood1, E. Johnstone1

1. Maternal & Fetal Health Research Centre, University of Manchester, Manchester, United Kingdom.


Pre-eclampsia and fetal growth restriction are common complications of pregnancy that can result in significant mortality and morbidity for both mother and child. They are both characterised by abnormal placental development including deficient invasion of the maternal endometrium by the extravillous trophoblast cells (EVT) of the placenta. Sphingosine-1-phosphate (S1P) is a signalling sphingolipid now recognised to play a role in a diverse array of fundamental biological processes including cell proliferation and migration; this occurs through 5 G protein-coupled S1P receptors (S1PR1-5)[1]. S1PR1 and S1PR3 have been shown to promote cell migration in vitro, whilst S1PR2 inhibits it. Our objective is to study the role of S1P and S1P receptors in EVT invasion of the maternal endometrium. Immunofluorescence using primary antibodies for S1PR1, 2 & 3 was carried out on Swan 71 cells and SGHPL-4 (two different human EVT derived cell lines) after culture under normoxic conditions. RT-PCR for S1PR1, 2 and 3 mRNA was carried after RNA extraction from Swan 71 cells. The results demonstrated evidence of expression of S1PR1, 2 & 3 protein expression in both Swan 71 and SGHPL-4 cell types (n=6). In addition RT-PCR on mRNA from Swan 71 cells confirmed the presence of S1PR1, 2, & 3 mRNA. Semi-quantitive analysis of immunofluoresence suggested relatively higher expression of S1PR3 than S1PR1 & 2. S1PR3 expression was also predominantly expressed at the loci of cell expansion, whereas S1PR1 & 2 were expressed uniformly through the cytoplasm. In conclusion, we have shown that S1P receptors are expressed on the external surface of the Swan 71 and SGHPL-4 EVT cell lines. Previous studies have suggested a predominance of S1PR3 leads to increased migration in the presence of S1P [2]. These data therefore imply that the S1P/S1PR system may regulate invasion and be a factor in the abnormal invasion seen in placental disease.

Where applicable, experiments conform with Society ethical requirements