期刊
JOURNAL OF CELL SCIENCE
卷 116, 期 15, 页码 3221-3226出版社
COMPANY OF BIOLOGISTS LTD
DOI: 10.1242/jcs.00627
关键词
membrane potential; PKC delta; superoxide; H+ channel; NADPH oxidase
类别
资金
- NIAID NIH HHS [AI50494-P1, AI34577-P4] Funding Source: Medline
Protein kinase C (PKC) activation in human eosinophils increases NADPH oxidase activity, which is associated with plasma membrane depolarization. In this study, membrane potential measurements of eosinophils stimulated with phorbol ester (phorbol 12-myristate 13-acetate; PMA) were made using a cell-permeable oxonol membrane potential indicator, diBAC(4)(3). Within 10 minutes after PMA stimulation, eosinophils depolarized from -32.9+/-5.7 mV to +17.3+/-1.8 mV. The time courses of depolarization and proton channel activation were virtually identical. Blocking the proton conductance with 250 muM ZnCl2 (+43.0+/-4.2 mV) or increasing the proton channel activation threshold by reducing the extracellular pH to 6.5 (+44.4+/-1.4 mV) increased depolarization compared with PNU alone. Additionally, the protein kinase C (PKC) delta-selective blocker, rottlerin, inhibited PMA-stimulated depolarization, indicating that PKCdelta was involved in regulating depolarization associated with eosinophil NADPH oxidase activity. Thus, the membrane depolarization that is associated with NADPH oxidase activation in eosinophils is sufficient to produce marked proton channel activation under physiological conditions.
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