Placental angiogenesis depends on AQP1 and AQP4, and the proper interaction with CAV-1

REPPETTI J.; REIS M.V.; CASAL J.J.; GORNALUSSE G.; DAMIANO A.E.; MARTINEZ N.

Mar del Plata

Congreso; Reunión anual de las sociedades de biociencias 2022 (SAIC, SAI, FAIC, SAFIS); 2022

PLACENTAL ANGIOGENESIS DEPENDS ON AQP1 AND AQP4, AND THE PROPER INTERACTION WITH CAV-1Julieta Reppetti1, Marcus V. Reis1, Juan J. Casal2, Germán Gornalusse3, Alicia E. Damiano1,4, Nora Martínez11 Laboratorio de Biología de la Reproducción. IFIBIO-Houssay (UBA-CONICET). Facultad de Medicina, Universidad de Buenos Aires. Buenos Aires, Argentina.2 IFIBIO Houssay (UBA-CONICET). Facultad de Medicina, Universidad de Buenos Aires. Buenos Aires, Argentina.3 Department of Medicine, University of Washington, Seattle, Washington, USA.4 Cátedra de Biología Celular y Molecular. Departamento de Ciencias Biológicas, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.Placental vasculature development involves the proliferation, migration, and differentiation of endothelial cells (EC). Caveolin-1 (Cav-1) is a constitutive protein of the caveola that interacts with numerous proteins. Aquaporins (AQPs) are transmembrane water channels, which move water in response to osmotic gradients. Emerging evidence shows that AQPs and Cav-1 are required for cell migration. We hypothesized that placental angiogenesis depends on the normal expression and function of AQPs that interact with Cav-1 in EC.Objective: To study the role of AQP1 and AQP4 in placental angiogenesis and their interaction with Cav-1.Materials and Methods: The macrovasculature cell line, EA.hy926 (ATCC®CRL-2922™), was used. AQP1, AQP4, and Cav-1 expressions were explored. In silico analysis was performed to investigate putative Cav-1 binding motifs in AQP1 and AQP4 proteins. Co-localization of these proteins was assessed by immunoprecipitation. AQPs were blocked with tetramethylammonium chloride (TEA), and Cav-1 and AQP1 were silenced by specific siRNA. Cell migration was evaluated by wound healing assay. Angiogenesis was evaluated by tube formation assay.Results: EA.hy926 cells expressed AQP1, AQP4, and Cav-1. In silico analysis identified a putative Cav-1-binding site in AQP1 and AQP4. Immunoprecipitation assay confirmed these results. Cav-1 silencing produced a significant reduction in cell migration (n=4; p