title: Human endometrial stromal cell rho GTPases have opposing roles in regulating focal adhesion turnover and embryo invasion in vitro.
abstract: Implantation of the embryo into the uterine compartment is a multistep event involving attachment of the embryo to the endometrial epithelia, followed by invasion of the embryo through the endometrial stroma. RHOA, RAC1, and CDC42 are members of the Rho GTPase family of proteins, which control cell functions such as cell migration and cytoskeletal reorganization. Herein, using a heterologous in vitro coculture model, we show that implantation of mouse blastocysts into human endometrial stromal cells (hESCs) is regulated by Rho GTPase activity in hESCs. Whereas iRNA-mediated silencing of RAC1 expression in hESCs led to inhibition of embryo implantation, silencing of either RHOA or CDC42 in hESCs promoted embryo implantation in coculture assays. Analysis of downstream signaling pathways demonstrated that RAC1 silencing was associated with decreased focal adhesion disassembly and resulted in large focal adhesion complexes in hESCs. In contrast, RHOA or CDC42 silencing resulted in perturbed focal adhesion assembly, leading to a decrease in the number of focal adhesions observed. Furthermore, inhibition of Rho signaling using a Rho kinase inhibitor, Y27632, led to decreased activation of protein tyrosine kinase 2 (PTK2, also called focal adhesion kinase) and decreased focal adhesion assembly. Importantly, perturbation of focal adhesion turnover in hESCs, mediated by PTK2 silencing, resulted in inhibition of embryo implantation into hESC monolayers. These findings suggest that Rho GTPase-PTK2-dependent remodeling of the endometrial stromal cell compartment may be critical for successful embryo implantation.
authors.affiliation: Nuffield Department of Obstetrics and Gynaecology, University of Oxford, Women's Centre, John Radcliffe Hospital, Oxford, United Kingdom.
authors.forename: Seema
authors.initials: S
authors.lastname: Grewal
authors.forename: Janet
authors.initials: J
authors.lastname: Carver
authors.forename: Anne J
authors.initials: AJ
authors.lastname: Ridley
authors.forename: Helen J
authors.initials: HJ
authors.lastname: Mardon
mesh_terms: D000328:Adult; D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002465:Cell Movement; D018920:Coculture Techniques; D010064:Embryo Implantation; D004717:Endometrium / Q000166:cytology / Q000201:enzymology*; D005260:Female; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism; D022001:Focal Adhesions / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D008875:Middle Aged; D034622:RNA Interference; D015398:Signal Transduction; D017154:Stromal Cells / Q000201:enzymology; D055815:Young Adult; D020741:rho GTP-Binding Proteins / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: D051417:Focal Adhesion Kinase 1; D054460:rho-Associated Kinases; D020741:rho GTP-Binding Proteins