Cloning and functional characterization of medaka TRPV4

Medaka, Oryzias latipes is distributed in fresh water of South Asia. To study roles of TRPV4 in osmosensing and adaption mechanism of medaka during changes in salinity environment, we isolated the cDNA for TRPV4 from medaka (olTRPV4) and characterized it. The electrophysiological analysis using Xenopus oocytes revealed that olTRPV4 can be activated by a TRPV4-specific agonist, GSK1016790A and acid at pH 5. Further, olTRPV4 was sensitive to 2-APB. Although warm temperatures activate mammalian TRPV4, olTRPV4 was activated by cold and hot stimulation. The threshold for cold activation was determined as 13.13 ± 0.60 °C, and the heat-activation threshold was 40.26 ± 0.25 °C. Further, when olTRPV4-expressing oocytes were stimulated by hypotonic solution, an apparent activation was observed. We further found that the expression of this hypotonic sensor, olTRPV4 was significantly down-regulated in gills but up-regulated in brains of sea water-acclimated medaka. Results demonstrated that olTRPV4 must function as an osmosensor and play important roles in adaption mechanism of medaka fish in various salinity environments.