DOI: 10.1152/ajplung.00015.2005. Effect of p47<sup><i>phox</i></sup>gene deletion on ROS production and oxygen sensing in mouse carotid body chemoreceptor cells

Effect of p47phoxgene deletion on ROS production and oxygen sensing in mouse carotid body chemoreceptor cells

10.1152/ajplung.00015.2005

Crossref journal-article

American Physiological Society

American Journal of Physiology-Lung Cellular and Molecular Physiology (24)

Abstract

Membrane potential in oxygen-sensitive type I cells in carotid body is controlled by diverse sets of voltage-dependent and -independent K+channels. Coupling of Po2to the open-closed state of channels may involve production of reactive oxygen species (ROS) by NADPH oxidase. One hypothesis suggests that ROS are produced in proportion to the prevailing Po2and a subset of K+channels closes as ROS levels decrease. We evaluated ROS levels in normal and p47phoxgene-deleted [NADPH oxidase knockout (KO)] type I cells using the ROS-sensitive dye dihydroethidium (DHE). In normal cells, hypoxia elicited an increase in ROS, which was blocked by the specific NADPH oxidase inhibitor 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF, 3 mM). KO type I cells did not respond to hypoxia, but the mitochondrial uncoupler azide (5 μM) elicited increased fluorescence in both normal and KO cells. Hypoxia had no effect on ROS production in sensory and sympathetic neurons. Methodological control experiments showed that stimulation of neutrophils with a cocktail containing the chemotactic peptide N-formyl-Met-Leu-Phe (1 μM), arachidonic acid (10 μM), and cytochalasin B (5 μg/ml) elicited a rapid increase in DHE fluorescence. This response was blocked by the NADPH oxidase inhibitor diphenyleneiodonium (10 μM). KO neutrophils did not respond; however, azide (5 μM) elicited a rapid increase in fluorescence. Physiological studies in type I cells demonstrated that hypoxia evoked an enhanced depression of K+current and increased intracellular Ca2+levels in KO vs. normal cells. Moreover, AEBSF potentiated hypoxia-induced increases in intracellular Ca2+and enhanced the depression of K+current in low O2. Our findings suggest that local compartmental increases in oxidase activity and ROS production inhibit the activity of type I cells by facilitating K+channel activity in hypoxia.

Bibliography

He, L., Dinger, B., Sanders, K., Hoidal, J., Obeso, A., Stensaas, L., Fidone, S., & Gonzalez, C. (2005). Effect of p47phoxgene deletion on ROS production and oxygen sensing in mouse carotid body chemoreceptor cells. American Journal of Physiology-Lung Cellular and Molecular Physiology, 289(6), L916âL924.

Authors 8

L. He (first)
B. Dinger (additional)
K. Sanders (additional)
J. Hoidal (additional)
A. Obeso (additional)
L. Stensaas (additional)
S. Fidone (additional)
C. Gonzalez (additional)

References 51 Referenced 44

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Dates

Type	When
Created	19 years, 9 months ago (Nov. 9, 2005, 3:33 p.m.)
Deposited	4 years, 1 month ago (July 19, 2021, 8:24 a.m.)
Indexed	4 months, 1 week ago (April 22, 2025, 3:13 a.m.)
Issued	19 years, 8 months ago (Dec. 1, 2005)
Published	19 years, 8 months ago (Dec. 1, 2005)
Published Print	19 years, 8 months ago (Dec. 1, 2005)

Funders 0

None

BibTeX

@article{He_2005, title={Effect of p47phoxgene deletion on ROS production and oxygen sensing in mouse carotid body chemoreceptor cells}, volume={289}, ISSN={1522-1504}, url={http://dx.doi.org/10.1152/ajplung.00015.2005}, DOI={10.1152/ajplung.00015.2005}, number={6}, journal={American Journal of Physiology-Lung Cellular and Molecular Physiology}, publisher={American Physiological Society}, author={He, L. and Dinger, B. and Sanders, K. and Hoidal, J. and Obeso, A. and Stensaas, L. and Fidone, S. and Gonzalez, C.}, year={2005}, month=dec, pages={L916–L924} }

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