ROLE OF THE CAVEOLAE AND AQP3 IN THE HUMAN PLACENTA DEVELOPMENT.

ALEJANDRA RECA; NORA MARTINEZ; JULIETA REPPETTI; NATALIA BELTRAMONE; ABRIL SEYAHIAN; ALICIA E. DAMIANO

Buenos Aires

Congreso; IV INTERNATIONAL CONGRESS OF TRANSLATIONAL MEDICINE; 2018

Introduction: Development of the human placenta is critical for a successful pregnancy. It requires that extravillous cytotrophoblast cells (EVTs) migrate, invade and transform maternal spiral arteries. Caveolae are membrane domains that compartmentalize intracellular signaling pathways to orchestrate different cellular events, such as cell migration and invasion. Caveolin-1 (Cav-1), the specific marker protein of caveolae, can bind to many membrane receptor proteins and concentrate these molecules in the caveolae. On the other hand, it was recently reported that aquaporin 3 (AQP3) is expressed in trophoblast cells during early gestation and its expression decreased dramatically in preeclamptic placentas. Although the caveolar domains and AQP3 are involved in the migration of different cell types, their importance and role during placentation remains unknown. Objective: To evaluate the role of caveolae and AQP3 in the migration and the invasion of EVT.Methods: Trophoblast cells (Swan 71 cell line) were cultured in complete DMEM-F12. AQP3 and Cav-1 expression was examined by Western blot, RT-PCR and immunofluorescence. Cells were treated with AQP3- siRNA to inhibit AQP3 and with 5 mM methyl-β-cyclodextrin (MCD) to disrupt caveolae. Migration was assessed by wound healing assay. Transwell invasion assay was also performed. Metalloproteinase activities were evaluated by gelatin gel zymography. Endovascular differentiation was evaluated by formation of tube-like structures in plates coated with Matrigel.Results: Cav-1 and AQP3 expression was found in Swan 71 cell line, and disruption of caveolae by MCD significantly decreased Cav-1 expression (n=5; p