Voltage gated proton channels modulate mitochondrial reactive oxygen species production by complex I in renal medullary thick ascending limb

Bansari Patel, Nadezhda N. Zheleznova, Sarah C. Ray, Jingping Sun, Allen W. Cowley, Paul M O'Connor

Research output: Contribution to journalArticle

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Abstract

Hv1 is a voltage-gated proton channel highly expressed in immune cells where, it acts to maintain NAD(P)H oxidase activity during the respiratory burst. We have recently reported that Hv1 is expressed in cells of the medullary thick ascending limb (mTAL) of the kidney and is critical to augment reactive oxygen species (ROS) production by this segment. While Hv1 is associated with NOX2 mediated ROS production in immune cells, the source of the Hv1 dependent ROS in mTAL remains unknown. In the current study, the rate of ROS formation was quantified in freshly isolated mTAL using dihydroethidium and ethidium fluorescence. Hv1 dependent ROS production was stimulated by increasing bath osmolality and ammonium chloride (NH 4 Cl) loading. Loss of either p67phox or NOX4 did not abolish the formation of ROS in mTAL. Hv1 was localized to mitochondria within mTAL, and the mitochondrial superoxide scavenger mitoTEMPOL reduced ROS formation. Rotenone significantly increased ROS formation and decreased mitochondrial membrane potential in mTAL from wild-type rats, while treatment with this inhibitor decreased ROS formation and increased mitochondrial membrane potential in mTAL from Hv1 −/− mutant rats. These data indicate that NADPH oxidase is not the primary source of Hv1 dependent ROS production in mTAL. Rather Hv1 localizes to the mitochondria in mTAL and modulates the formation of ROS by complex I. These data provide a potential explanation for the effects of Hv1 on ROS production in cells independent of its contribution to maintenance of cell membrane potential and intracellular pH.

Original languageEnglish (US)
Article number101191
JournalRedox Biology
DOIs
StatePublished - Jan 1 2019

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Protons
Reactive Oxygen Species
Extremities
Kidney
Electric potential
Mitochondria
Mitochondrial Membrane Potential
NADPH Oxidase
Rats
Membranes
Rotenone
Ammonium Chloride
Ethidium
Respiratory Burst
Cell membranes
Baths
Superoxides
Membrane Potentials
Osmolar Concentration
Fluorescence

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry

Cite this

Voltage gated proton channels modulate mitochondrial reactive oxygen species production by complex I in renal medullary thick ascending limb. / Patel, Bansari; Zheleznova, Nadezhda N.; Ray, Sarah C.; Sun, Jingping; Cowley, Allen W.; O'Connor, Paul M.

In: Redox Biology, 01.01.2019.

Research output: Contribution to journalArticle

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AU - Cowley, Allen W.

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