Regulation of eosinophil membrane depolarization during NADPH oxidase activation

Jennifer L. Bankers-Fulbright, Gerald J. Gleich, Gail M. Kephart, Hirohito Kita, Scott M. O'Grady

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

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, diBAC4(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 μM 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 PMA alone. Additionally, the protein kinase C (PKC) δ-selective blocker, rottlerin, inhibited PMA-stimulated depolarization, indicating that PKCδ 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.

Original languageEnglish (US)
Pages (from-to)3221-3226
Number of pages6
JournalJournal of Cell Science
Volume116
Issue number15
DOIs
StatePublished - Aug 1 2003

Fingerprint

NADPH Oxidase
Eosinophils
Protons
Membranes
Protein Kinase C
Membrane Potentials
Phorbol Esters
Acetates
Cell Membrane

Keywords

  • H channel
  • Membrane potential
  • NADPH oxidase
  • PKCδ
  • Superoxide

ASJC Scopus subject areas

  • Cell Biology

Cite this

Bankers-Fulbright, J. L., Gleich, G. J., Kephart, G. M., Kita, H., & O'Grady, S. M. (2003). Regulation of eosinophil membrane depolarization during NADPH oxidase activation. Journal of Cell Science, 116(15), 3221-3226. https://doi.org/10.1242/jcs.00627

Regulation of eosinophil membrane depolarization during NADPH oxidase activation. / Bankers-Fulbright, Jennifer L.; Gleich, Gerald J.; Kephart, Gail M.; Kita, Hirohito; O'Grady, Scott M.

In: Journal of Cell Science, Vol. 116, No. 15, 01.08.2003, p. 3221-3226.

Research output: Contribution to journalArticle

Bankers-Fulbright, JL, Gleich, GJ, Kephart, GM, Kita, H & O'Grady, SM 2003, 'Regulation of eosinophil membrane depolarization during NADPH oxidase activation', Journal of Cell Science, vol. 116, no. 15, pp. 3221-3226. https://doi.org/10.1242/jcs.00627
Bankers-Fulbright, Jennifer L. ; Gleich, Gerald J. ; Kephart, Gail M. ; Kita, Hirohito ; O'Grady, Scott M. / Regulation of eosinophil membrane depolarization during NADPH oxidase activation. In: Journal of Cell Science. 2003 ; Vol. 116, No. 15. pp. 3221-3226.
@article{affb34f5daca4200a242d58cbaf2876f,
title = "Regulation of eosinophil membrane depolarization during NADPH oxidase activation",
abstract = "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, diBAC4(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 μM 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 PMA alone. Additionally, the protein kinase C (PKC) δ-selective blocker, rottlerin, inhibited PMA-stimulated depolarization, indicating that PKCδ 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.",
keywords = "H channel, Membrane potential, NADPH oxidase, PKCδ, Superoxide",
author = "Bankers-Fulbright, {Jennifer L.} and Gleich, {Gerald J.} and Kephart, {Gail M.} and Hirohito Kita and O'Grady, {Scott M.}",
year = "2003",
month = "8",
day = "1",
doi = "10.1242/jcs.00627",
language = "English (US)",
volume = "116",
pages = "3221--3226",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "15",

}

TY - JOUR

T1 - Regulation of eosinophil membrane depolarization during NADPH oxidase activation

AU - Bankers-Fulbright, Jennifer L.

AU - Gleich, Gerald J.

AU - Kephart, Gail M.

AU - Kita, Hirohito

AU - O'Grady, Scott M.

PY - 2003/8/1

Y1 - 2003/8/1

N2 - 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, diBAC4(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 μM 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 PMA alone. Additionally, the protein kinase C (PKC) δ-selective blocker, rottlerin, inhibited PMA-stimulated depolarization, indicating that PKCδ 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.

AB - 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, diBAC4(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 μM 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 PMA alone. Additionally, the protein kinase C (PKC) δ-selective blocker, rottlerin, inhibited PMA-stimulated depolarization, indicating that PKCδ 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.

KW - H channel

KW - Membrane potential

KW - NADPH oxidase

KW - PKCδ

KW - Superoxide

UR - http://www.scopus.com/inward/record.url?scp=0041885139&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0041885139&partnerID=8YFLogxK

U2 - 10.1242/jcs.00627

DO - 10.1242/jcs.00627

M3 - Article

C2 - 12829741

AN - SCOPUS:0041885139

VL - 116

SP - 3221

EP - 3226

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 15

ER -