Role of Aquaporin-2 and TRPV4 In Renal Cell Migration.

ALEJANDRO PIZZONI; VALERIA RIVAROLA; GISELA DI GIUSTO; NATALIA BELTRAMONE; CLAUDIA CAPURRO; MARIA FLORENCIA RIVERA; PAULA FORD

Congreso; . Reunión Conjunta de Biociencias SAIC-SAI-SAFIS (2020); 2020

Role of Aquaporin-2 and TRPV4 in renal cell migrationWe have previously shown that Aquaporin-2 (AQP2) promotes renal cell migration. Thispromigratory effect is due, at least in part, to the modulation of Na+/H+ exchanger (NHE1)activity, responsible for lamellipodia alkalinization, which would generate the appropriatemicroenvironment for actin and focal adhesions dynamics. Taking into account that wehave recently demonstrated a physical and functional interaction between AQP2 and thecalcium channel TRPV4, and that NHE1 activity is modulated by calcium, we propose toinvestigate the contribution of TRPV4 in the AQP2-dependent renal cell migration. Theexperimental procedures were performed using two renal cortical collecting duct cellmodels: one WT not expressing AQPs and another one expressing AQP2. Usingimmunofluorescence assays, we first determined TRPV4 expression in lamellipodia ofmigrating cells. Then, we investigated TRPV4 participation in collective cell migrationthrough wound healing assays in the presence of TRPV4 specific activator(GSK1016790A) and inhibitor (HC-067047). Finally, we characterize focal adhesioncomplexes by revealing Vinculin, a protein described as mechanosensor of theseadhesive sites. Our results showed that TRPV4 is present in lamellipodia of both celltypes, but AQP2-expressing cells have a higher intensity ratio per area analyzed (WT:794±84, n=32; AQP2: 1371±79, n=74; ***p