Natanson-Yaron Shira, Anteby Eyal Y, Greenfield Caryn, Goldman-Wohl Debra, Hamani Yaron, Hochner-Celnikier Drorit, Yagel Simcha
Department of Obstretrics and Gynecology, Hadassah University Hospital Mount Scopus, PO Box 24035, Mount Scopus, Jerusalem 91240, Israel.
Mol Hum Reprod. 2007 Jul;13(7):511-9. doi: 10.1093/molehr/gam034. Epub 2007 May 12.
Branching morphogenesis (BM) of the chorionic villous tree is a crucial component of early placental formation. Fibroblast growth factors (FGFs), their receptor tyrosine kinase (RTK) and negative regulators like Sprouty (Spry) proteins are pivotal factors in the development of diverse branching organ systems. The aim of this study was to examine the effect of FGF10 and Sprouty 2 on BM of the chorionic villi in vitro. Villous explants of first trimester placentas were cultured and their outgrowths were monitored. The effect of FGF10 was tested on matrigel migration/invasion assay, collagenolytic activity of single cell trophoblasts and on villous explants outgrowths. siRNA of Spry2 was used to reduce its expression and to investigate the role of Sprouty 2 in villous explants outgrowths. Quantitative RT-PCR and immunohistochemistry were performed to determine Sprouty 2 and HLA-G (a marker of invasion) expression. FGF 10 stimulated by 8-fold the migration/invasion of single cell trophoblast enhanced their collagenolytic activity. Reduction of Spry2 expression in villous explants showed a marked increase in villous outgrowths. This was accompanied by enhanced staining for HLA-G and by the reduction of Spry2 expression that was confirmed by immunohistochemistry and by quantitative RT-PCR. We conclude that trophoblast outgrowth and invasion (part of placental villi sprouting) at the fetal maternal interface is in part under delicate control of FGF 10 and Sprouty 2. FGF 10 promotes invasion and outgrowth of trophoblasts. In addition, it increases Spry2 expression, which attenuates trophoblast sprouting.
绒毛膜绒毛树的分支形态发生（BM）是早期胎盘形成的关键组成部分。成纤维细胞生长因子（FGFs）、其受体酪氨酸激酶（RTK）以及像Sprouty（Spry）蛋白这样的负调节因子是多种分支器官系统发育中的关键因素。本研究的目的是在体外检测FGF10和Sprouty 2对绒毛膜绒毛BM的影响。培养孕早期胎盘的绒毛外植体并监测其生长。在基质胶迁移/侵袭试验、单细胞滋养层细胞的胶原olytic活性以及绒毛外植体生长方面测试FGF10的作用。使用Spry2的小干扰RNA（siRNA）降低其表达，并研究Sprouty 2在绒毛外植体生长中的作用。进行定量逆转录聚合酶链反应（RT-PCR）和免疫组织化学以确定Sprouty 2和HLA-G（侵袭标志物）的表达。FGF 10刺激单细胞滋养层细胞的迁移/侵袭增加8倍，增强了它们的胶原olytic活性。绒毛外植体中Spry2表达的降低显示绒毛生长显著增加。这伴随着HLA-G染色增强以及Spry2表达降低，免疫组织化学和定量RT-PCR证实了这一点。我们得出结论，胎儿-母体界面处的滋养层生长和侵袭（胎盘绒毛发芽的一部分）部分受FGF 10和Sprouty 2的精细控制。FGF 10促进滋养层细胞的侵袭和生长。此外，它增加Spry2表达，从而减弱滋养层发芽。
