The Role Of Protein Kinase C Alpha In Aquaporin-2 Trafficking

Objective: Protein kinase Cα(PKCα) is recognized for its role in urine concentration, but the detail mechanism is not very clear. Here we investigated the role of protein kinase Cα(PKCα) in aquaporin 2(AQP2) trafficking and whether cytoskeleton (CSK) is involved in the process.Methods: AQP2-GFP plasmid was stably transfected into mIMCD₃ cells, Western blot and immunofluoresence was used to detect the transfection efficiency; AQP2-mIMCD₃ cells were transfected with PKCαshRNA, A25E and scramble shRNA plasmid respectively for 48hours, and then treated with DdAVP for 30min. Cells were harvested at 0min, 30min after DdAVP treatment, and DdAVP treated 30min then removed after 2h by washout, distribution of AQP2 and microtubule were observed by confocal microscope.Results: AQP2-mIMCD₃ cells were successfully established and the transfection efficiency was confirmed by immunolofluorescence microscope and Western blot. Moreover, as compared to PKCαactivation plasmid group and control plasmid group, after being transfected with PKCαshRNA plasmid, DdAVP stimulated AQP2 trafficking was abolished in AQP2-mIMCD₃ cell, the latter was accompanied by an unchanged microtubule distribution. Conclusion: PKCαplays a role in AQP2 trafficking, which probably through mediates the dynamics feature and distribution of microtubule in mIMCD₃ cells.