An intact caveolar structure is necessary for the proper formation of placental microvasculature.

REPPETTI J.; SIERRA M.; ANUD C.; ERLEJMAN A.; DAMIANO A. E.; MARTÍNEZ N.

Rosario

Congreso; REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE FISIOLOGÍA; 2019

SAFIS

An intact caveolar structure is necessary for the proper formation of placental microvasculatureReppetti Julieta1, Sierra Matías N.2, Anud Carolina2, Erlejman Alejandra3, Damiano Alicia E.1,2, Martínez Nora1.1 Laboratorio de Biología de la Reproducción. Instituto de Fisiología y Biofísica Bernardo Houssay (IFIBIO Houssay) UBA-CONICET. Buenos Aires, Argentina.2 Cátedra de Biología Celular y Molecular. Departamento de Ciencias Biológicas, Facultad de Farmacia y Bioquímica, UBA. Buenos Aires, Argentina.3 Laboratorio de Biología Molecular y Celular. Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, UBA. IQUIBICEN- CONICET. Buenos Aires, ArgentinaIntroduction: A proper development of the placental vasculature is necessary for a successful pregnancy. Two types of endothelial cells form the human placenta: the microvascular endothelial cells which comprise the fetal capillaries of chorionic villi and are involved in the oxygen and nutrients exchange, and the macrovascular endothelial cells which are the large conduit vessels of the umbilical cord. These cells differ in morphology, gene expression and function.Caveolae are a type of lipid rafts, coated with caveolin-1 (Cav-1) protein. On the other hand, Aquaporin-1 (AQP1) is a transmembrane water channel which moves water in response to osmotic gradients. Recently, it was suggested that AQPs and Cav-1 are also indispensable for efficient cell migration. Objective: To evaluate the role of caveolae in the formation of placental microvasculature and its interaction with AQP1.Materials and Methods: This study was approved by the ethics committee of the Hospital Nacional Prof. Dr. A. Posadas. Placental microvascular endothelial cells (hPMEC) and EA.hy926 cell line (ATCC® CRL-2922?) were used. Gene expression of Cav-1 and AQP1 was evaluated by RT-PCR. Protein expression and localization were studied by Western Blot and immunofluorescence. Co-localization of Cav-1 with AQP1 was assessed by immunoprecipitation assay. Cells were treated with 5 mM methyl-β-cyclodextrin (MβCD) to disrupt caveolae and with Tetraethylammonium chloride (TEA) to block AQP1. Cell migration was evaluated by wound healing assay. Results: Cav-1 and AQP1 co-localized in the cell membrane of placental endothelial cells. MβCD significantly reduced cell migration in EA.hy926 cells (n=4; p