AQUAPORINS AND CAVEOLIN-1 IN PLACENTAL VASCULAR DEVELOPMENT

JULIETA REPPETTI; GERMÁN GORNALUSSE; TOMÁS ETCHEVERRY; ALICIA DAMIANO; NORA A MARTÍNEZ

Simposio; 9th Latin American Symposium on Maternal-Fetal Interaction and Placenta (IX SLIMP); 2022

Objective: Proper development of the placental macrovasculature and microvasculature is necessary to ensure a successful pregnancy. We have previously demonstrated the expression of aquaporin-1 (AQP1) and caveolin-1 (Cav-1) in placental endothelial cells, and since they fulfill different functions, this study aimed to compare the differential behavior of the macrovascular and microvascular cells, focusing on the role of the caveolae and AQPs.Methodology: Primary culture of human placental microvascular endothelial cells (hPMEC) was cultured in complete M199. This study was approved by the ethics committee of the Hospital Nacional Prof. Dr. A. Posadas. Macrovasculature cell line EA.hy926 (ATCC® CRL-2922™) was cultured in complete DMEM/F-12.Gene expression of Cav-1, AQP1, and AQP4 was evaluated by RT-qPCR. Protein expression and localization were studied by Western Blot and immunofluorescence. Cells were treated with pharmacological inhibitors: tetramethylammonium chloride (TEA) to block AQPs, methyl-β-cyclodextrin (MβCD) to disrupt caveolae and with specific siRNAs to silence protein expression, and cell migration was evaluated by the wound healing assay.Results: hPMEC and EA.hy926 cells express Cav-1, AQP1, and AQP4. AQP4 levels were higher in EA.hy926 than in hPMEC. In both cases, the inhibition of Cav-1 or the disruption of the caveola produced a dramatic reduction in cell migration (n=4; p