Study of the role of caveolae/ Caveolin-1 in the early development of the human placenta

REPPETTI JULIETA; SEYAHIAN ABRIL; RECA ALEJANDRA; SIERRA MATÍAS N; DAMIANO ALICIA E; MARTÍNEZ NORA

Workshop; Workshop IFIBIO; 2018

Development of the human placenta is critical for a successful pregnancy. It requires that extravillous cytotrophoblast cells (EVTs) migrate and invade the uterine decidua and adopt an endothelial phenotype to modify the utero-placental 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 many membrane receptor proteins and concentrate these molecules in the caveolae. However, the importance of caveolae during placentation remains unknown.Our aim was to investigate the role of caveolae present in the human trophoblast in the processes of proliferation, migration, invasion and endovascular differentiation.Swan 71 cell line was treated with 5 mM methyl-β-cyclodextrin (MβCD) to disrupt caveolae. Cav-1 expression was examined by Western blot, RT-PCR (sense 5?-TCTCTACACCGTTCCCATCC-3? and antisense 5?-CACAGACGGTGTGGACGTAG-3?) and immunofluorescence. Cell migration was assessed by wound healing assay. Transwell invasion assay was also performed. Metalloproteinase activities were evaluated by gelatin zymography. Endovascular differentiation was evaluated by formation of tube-like structures in plates coated with Matrigel®.Disruption of caveolae significantly decreased Cav-1 expression (n=3; p