Identification of a neuronal nitric oxide synthase in isolated cardiac mitochondria using electrochemical detection
Open accessAbstract
Mitochondrial nitric oxide synthase (mtNOS), its cellular NOS isoform, and the effects of mitochondrially produced NO on bioenergetics have been controversial since mtNOS was first proposed in 1995. Here we functionally demonstrate the presence of a NOS in cardiac mitochondria. This was accomplished by direct porphyrinic microsensor measurement of Ca 2+ -dependent NO production in individual mitochondria isolated from wild-type mouse hearts. This NO production could be inhibited by NOS antagonists or protonophore collapse of the mitochondrial membrane potential. The similarity of mtNOS to the neuronal isoform was deduced by the absence of NO production in the mitochondria of knockout mice for the neuronal, but not the endothelial or inducible, isoforms. The effects of mitochondrially produced NO on bioenergetics were studied in intact cardiomyocytes isolated from dystrophin-deficient ( mdx ) mice. mdx cardiomyocytes are also deficient in cellular endothelial NOS, but overexpress mtNOS, which allowed us to study the mitochondrial enzyme in intact cells free of its cytosolic counterpart. In these cardiomyocytes, which produce NO beat-to-beat, inhibition of mtNOS increased myocyte shortening by approximately one-fourth. Beat-to-beat NO production and altered shortening by NOS inhibition were not observed in wild-type cells. A plausible mechanism for the reversible NO inhibition of contractility in these cells involves the reaction of NO with cytochrome c oxidase. This suggests a modulatory role for NO in oxidative phosphorylation and, in turn, myocardial contractility.
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Microsensor measurements of single isolated mitochondria provide some evidence for NO production in mitochondria.
“microsensors were used to monitor the release of NO from single isolated mitochondria, providing some evidence for NO production in mitochondria (208).”
From Redox regulation of mitochondrial function (2012) · cites this paper as [208]
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- Authors
- Kanai, A. J., Pearce, L. L., Clemens, P. R., Birder, L. A., VanBibber, M. M., Choi, S.-Y., de Groat, W. C., & Peterson, J.
- Affiliations
- University of Pittsburgh (US)
- University of California, Los Angeles (US)
- Departments of Medicine, Laboratory of Epithelial Cell Biology, Renal-Electrolyte Division, Neurology, and Pharmacology, University of Pittsburgh, Pittsburgh, PA 15261; Neurology Service, Veterans Affairs Pittsburgh Healthcare System, Pittsburgh, PA 15240; and Department of Chemistry, Carnegie Mellon University, Pittsburgh, PA 15213
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2001-11-20
- Volume
- 98
- Issue
- 24
- Pages
- 14126-14131
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC61179
- OpenAlex
- W2143445854
- Cited by
- 341
- References
- 53
- Citations by year
- 2026: 3 · 2025: 1 · 2024: 4 · 2023: 3 · 2022: 4 · 2021: 9 · 2020: 11 · 2019: 6 · 2018: 6 · 2017: 6 · 2016: 6 · 2015: 8 · 2014: 11 · 2013: 9 · 2012: 12
- FWCI
- 9.08
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Heart, Lung, and Blood Institute (HL-61411, R01HL061411)
- National Institute of Diabetes and Digestive and Kidney Diseases (R37DK054824, DK-54824, R01DK054824)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Health Sciences › Medicine › Pathology and Forensic Medicine › Cardiac Ischemia and Reperfusion
- Keywords
- mitochondrial nitric oxide synthase, neuronal nitric oxide synthase, heart mitochondria, nitric oxide production, mitochondrial membrane potential, NOS isoforms, mdx mice, cardiomyocyte contractility, cytochrome c oxidase, oxidative phosphorylation, myocardial bioenergetics
- MeSH terms
- Animals, Calcium, Electron Transport Complex IV, Electrochemistry, Mitochondria, Heart, Nitric Oxide, Superoxides, Dystrophin, Mice, Knockout, Nitric Oxide Synthase, Calcium Signaling, Mice, Nitric Oxide Synthase Type I
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