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Do mitochondria make nitric oxide? no?

Yvonne Mei Sian Tay1, Kok Seong Lim, Fwu-Shan Sheu

  • 1Department of Biochemistry, Faculty of Medicine, National University of Singapore, MD 7 #02-07, 8 Medical Drive, Singapore 117597, Singapore.

Free Radical Research
|September 7, 2004
PubMed
Summary
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This study found no evidence of nitric oxide (NO) production in rat liver mitochondria, challenging previous claims of mitochondrial nitric oxide synthase (NOS) activity and its impact on respiration.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Mitochondrial Physiology

Background:

  • Previous research suggested mitochondria possess nitric oxide synthase (NOS) and produce nitric oxide (NO) affecting respiration.
  • The existence and functional significance of mitochondrial NOS remain debated.

Purpose of the Study:

  • To investigate the presence and activity of nitric oxide synthase (NOS) in intact rat liver mitochondria.
  • To determine if mitochondria produce detectable levels of nitric oxide (NO) that influence respiratory function.

Main Methods:

  • Assessed the effect of L-arginine and L-NMMA on mitochondrial respiration rates (State 3 and State 4).
  • Utilized oxymyoglobin oxidation assay and electrochemical NO electrodes to detect mitochondrial NO production.
  • Investigated NO consumption by succinate-supplemented mitochondria and NO detection using the Griess assay.

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Main Results:

  • Neither L-arginine nor L-NMMA altered mitochondrial respiratory rates.
  • No mitochondrial NO production was detected using oxymyoglobin or electrochemical methods.
  • The Griess assay detected apparent NO production, identified as an artifact.
  • Exogenously added NO was detectable, but mitochondria consumed it, especially when supplemented with succinate.

Conclusions:

  • Rat liver mitochondria do not produce detectable levels of nitric oxide (NO) under the tested conditions.
  • The claimed mitochondrial nitric oxide synthase (NOS) activity and its impact on respiration are not supported by these findings.
  • Artifacts may explain previous positive results for mitochondrial NO production.