FOCAL ADHESION ASSEMBLY/DISASSEMBLY AND TRPV4 PARTICIPATION IN AQP2-DEPENDENT RENAL CELL MIGRATION

CAPURRO CLAUDIA; RIVAROLA VALERIA; DI GIUSTO GISELA; NATALIA BELTRAMONE; WHITE ALAN; DI GIUSTO GISELA; NATALIA BELTRAMONE; WHITE ALAN; FORD PAULA; PIZZONI ALEJANDRO; FORD PAULA; PIZZONI ALEJANDRO; CAPURRO CLAUDIA; RIVAROLA VALERIA

Buenos Aires

Congreso; Reunión Conjunta de Sociedades de Biociencias; 2017

SAFIS

We have previously demonstrated that Aquaporin 2 (AQP2) promotesrenal cell migration. It is well known that migration is a processthat requires continuous turnover of focal adhesions (FAs) andit was shown that AQP2 interacts with proteins forming FAs, so theaim of the present work was to study the dynamics of FAs in renalcells expressing AQP2. Moreover, since Ca2+ signaling is implicatedin FAs turnover and results from our laboratory showed a differentialactivation of the Ca2+ channel TRPV4 influenced by AQP2, wealso investigated if TRPV4 participates in the AQP2-enhanced renalcell migration. For experimental procedures two renal cell lineswere used: WT-RCCD1 (not expressing AQPs) and AQP2-RCCD1(transfected with AQP2). Immunofluorescence studies with paxillinwere performed in scratched monolayers to visualize FAs. Imageswere taken with a confocal microscope, deconvolved and processedfor making FAs area measurements. Cell migration in presence ofTRPV4 agonists 4α-PDD (10 mM) and GSK1016790A (3 nM) wasinvestigated with Wound Healing assay. Images were taken at differenttimes and results were expressed as % of wound closure.Immunofluorescence studies showed punctuated paxillin labelingindicating FAs in the leading edge of migrating RCCD1 cells. TheseFAs were found to be small in AQP2-RCCD1 (1.80±0.19 mm2, n=85)than in WT-RCCD1 (5.87±1.04 mm2, n=120, p