Am J Physiol Cell Physiol  AJP: Regulatory, Integrative and Comparative Physiology
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Am J Physiol Cell Physiol 293: C390-C400, 2007. First published May 30, 2007; doi:10.1152/ajpcell.00104.2007
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MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

Activation and inactivation of the volume-sensitive taurine leak pathway in NIH3T3 fibroblasts and Ehrlich Lettre ascites cells

Ian Henry Lambert

Department of Molecular Biology, University of Copenhagen, Copenhagen, Denmark

Submitted 15 March 2007 ; accepted in final form 29 April 2007

Hypotonic exposure provokes the mobilization of arachidonic acid, production of ROS, and a transient increase in taurine release in Ehrlich Lettre cells. The taurine release is potentiated by H2O2 and the tyrosine phosphatase inhibitor vanadate and reduced by the phospholipase A2 (PLA2) inhibitors bromoenol lactone (BEL) and manoalide, the 5-lipoxygenase (5-LO) inhibitor ETH-615139, the NADPH oxidase inhibitor diphenyl iodonium (DPI), and antioxidants. Thus, swelling-induced taurine efflux in Ehrlich Lettre cells involves Ca2+-independent (iPLA2)/secretory PLA2 (sPLA2) plus 5-LO activity and modulation by ROS. Vanadate and H2O2 stimulate arachidonic acid mobilization and vanadate potentiates ROS production in Ehrlich Lettre cells and NIH3T3 fibroblasts under hypotonic conditions. However, vanadate-induced potentiation of the volume-sensitive taurine efflux is, in both cell types, impaired in the presence of BEL and DPI and following restoration of the cell volume. Thus, potentiation of the volume-sensitive taurine efflux pathway following inhibition of tyrosine phosphatase activity reflects increased arachidonic acid mobilization and ROS production for downstream signaling. Vanadate delays the inactivation of volume-sensitive taurine efflux in NIH3T3 cells, and this delay is impaired in the presence of DPI. Vanadate has no effect on the inactivation of swelling-induced taurine efflux in Ehrlich Lettre cells. It is suggested that increased tyrosine phosphorylation of regulatory components of NADPH oxidase leads to increased ROS production and a subsequent delay in inactivation of the volume-sensitive taurine efflux pathway and that NADPH oxidase or antioxidative capacity differ between NIH3T3 and Ehrlich Lettre cells.

organic osmolytes; reactive oxygen species; vanadate; H2O2; tyrosine phosphatases; arachidonic acid mobilization



Address for reprint requests and other correspondence: I. H. Lambert, Dept. of Molecular Biology, The August Krogh Bldg., Universitetsparken 13, Copenhagen Ø DK-2100, Denmark (e-mail: ihlambert{at}aki.ku.dk)




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