ROLE OF CAVEOLAE IN THE FORMATION OF THE PLACENTAL MICROVASCULATURE AND ITS INTERACTION WITH AQP1 AND VEGFR2.

JULIETA REPPETTI; MATÍAS SIERRA; CARLOS ESCUDERO; ALICIA DAMIANO; NORA MARTÍNEZ

CABA

Congreso; IFPA 2019-VIII SLIMP; 2019

Objective: To evaluate the role of caveolae in the formation of the placental microvasculature and its interaction with AQP1 and VEGFR2.Materials and Methods: Primary culture of 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. Gene expression of Cav-1, AQP1 and VEGFR2 in macrovascular and microvascular endothelial cells was evaluated by RT-PCR. Protein expression and localization were studied by Western Blot and immunofluorescence. Co-localization of Cav-1 with AQP1 and VEGFR2 was assessed by immunoprecipitation assay. Cells were treated with 5 mM methyl-β-cyclodextrin (MβCD) to disrupt caveolae and cell migration was evaluated by the wound healing assay. Macrovasculature cell line EA.hy926 (ATCC® CRL-2922 ?) derived from the fusion of human umbilical vein macrovascular endothelial cells (HUVEC) with the human cell line A549 were used as controls. Results: hPMEC cells express Cav-1, AQP1 and VEGFR2. All these proteins were localized in the cell membrane of hPMEC and AQP1 and VEGFR2 co-localize with Cav-1. Treatment with MβCD significantly reduced cell migration (p