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Volume-Sensitive Chloride Channels Do Not Mediate Activation-Induced Chloride Efflux in Human Neutrophils¹

Patricia Perez-Cornejo^*,, Jorge Arreola^,, Foon-Yee Law^*, Joanne B. Schultz^* and Philip A. Knauf^2,*

^* Department of Biochemistry and Biophysics, School of Medicine, and Center for Oral Biology, Aab Institute of Biomedical Sciences, University of Rochester, Rochester, NY 14642; and Facultad de Medicina and Instituto de Fisica Universidad Autonoma de San Luis Potosi, San Luis Potosi, Mexico

Many agents that activate neutrophils, enabling them to adhere to venular walls at sites of inflammation, cause a rapid Cl^– efflux. This Cl^– efflux and the increase in the number and affinity of ₂ integrin surface adhesion molecules (up-regulation) are all inhibited by ethacrynic acid and certain aminomethyl phenols. The effectiveness of the latter compounds correlates with their inhibition of swelling-activated Cl^– channels (I_Clvol), suggesting that I_Clvol mediates the activator-induced Cl^– efflux. To test this hypothesis, we used whole-cell patch clamp in hypotonic media to examine the effects of inhibitors of up-regulation on I_Clvol in neutrophils and promyelocytic leukemic HL-60 cells. Both the channel blocker 5-nitro-2-(3-phenylpropylamino)benzoic acid and [3-methyl-1-p-sulfophenyl-5-pyrazolone-(4)]-[1,3-dibutylbarbituric acid]-pentamethine oxonol (WW781), a nonpenetrating oxonol, inhibited I_Clvol at concentrations similar to those that inhibit ₂ integrin up-regulation. However, ethacrynic acid, at the same concentration that inhibits activator-induced Cl^– efflux and up-regulation, had no effect on I_Clvol and swelling-activated Cl^– efflux, providing evidence against the involvement of I_Clvol in the activator-induced Cl^– efflux.

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