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1 Department of Biochemistry, Erasmus University Medical Center, Rotterdam, The Netherlands
2 Department of Psychiatry, Erasmus University Medical Center, Rotterdam, The Netherlands
* To whom correspondence should be addressed. E-mail: b.tilly{at}ErasmusMC.nl.
Human Intestine 407 cells respond to osmotic cell swelling by the activation of Cl- and K+ selective ionic channels, as well as by stimulating an organic osmolyte release pathway, readily permeable to taurine and phosphocholine. Unlike the activation of Volume Regulated Anion Channels (VRAC's), activation of the organic osmolyte release pathway shows a lag time of approx 30 - 60s and its activity persists for at least 8-12 minutes. In contrast to VRAC activation, stimulation of organic osmolyte release did not require protein tyrosine phosphorylation, active p21Rho or PtdIns-3-kinase activity and was insensitive to Cl- channel blockers. Treatment of the cells with putative Organic Anion Transporter (OAT) inhibitors reduced the release of taurine only partially or was found to be ineffective. The efflux was blocked by a subclass of Organic Cation Transporter (OCT) inhibitors (cyanine-863 and decynium-22), but not by other OCT inhibitors (cimetidine, quinine and verapamil). Brief treatment of the cells with phorbol esters potentiated the cell swelling-induced taurine efflux, whereas addition of the protein kinase C inhibitor GF109203X largely inhibited the response, suggesting that protein kinase C is involved. Raising intracellular Ca2+, using A23187 or Ca2+ mobilizing hormones, however, did not affect the magnitude of the response. Taken together, the results indicate that the hypotonicity-induced efflux of organic osmolytes is independent of VRAC and involves a PKC dependent step.
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