N-acetylcysteine and cysteamine bitartrate prevent azide-induced neuromuscular decompensation by restoring glutathione balance in two novel surf1-/- zebrafish deletion models of Leigh syndrome

Hum Mol Genet. 2023 Jun 5;32(12):1988-2004. doi: 10.1093/hmg/ddad031.


SURF1 deficiency (OMIM # 220110) causes Leigh syndrome (LS, OMIM # 256000), a mitochondrial disorder typified by stress-induced metabolic strokes, neurodevelopmental regression and progressive multisystem dysfunction. Here, we describe two novel surf1-/- zebrafish knockout models generated by CRISPR/Cas9 technology. While gross larval morphology, fertility, and survival into adulthood appeared unaffected, surf1-/- mutants manifested adult-onset ocular anomalies and decreased swimming activity, as well as classical biochemical hallmarks of human SURF1 disease, including reduced complex IV expression and enzymatic activity and increased tissue lactate. surf1-/- larvae also demonstrated oxidative stress and stressor hypersensitivity to the complex IV inhibitor, azide, which exacerbated their complex IV deficiency, reduced supercomplex formation, and induced acute neurodegeneration typical of LS including brain death, impaired neuromuscular responses, reduced swimming activity, and absent heartrate. Remarkably, prophylactic treatment of surf1-/- larvae with either cysteamine bitartrate or N-acetylcysteine, but not other antioxidants, significantly improved animal resiliency to stressor-induced brain death, swimming and neuromuscular dysfunction, and loss of heartbeat. Mechanistic analyses demonstrated cysteamine bitartrate pretreatment did not improve complex IV deficiency, ATP deficiency, or increased tissue lactate but did reduce oxidative stress and restore glutathione balance in surf1-/- animals. Overall, two novel surf1-/- zebrafish models recapitulate the gross neurodegenerative and biochemical hallmarks of LS, including azide stressor hypersensitivity that was associated with glutathione deficiency and ameliorated by cysteamine bitartrate or N-acetylcysteine therapy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcysteine
  • Adult
  • Animals
  • Azides / metabolism
  • Brain Death
  • Cysteamine / pharmacology
  • Cytochrome-c Oxidase Deficiency*
  • Electron Transport Complex IV / genetics
  • Electron Transport Complex IV / metabolism
  • Glutathione / metabolism
  • Humans
  • Lactates
  • Leigh Disease* / drug therapy
  • Leigh Disease* / genetics
  • Leigh Disease* / metabolism
  • Membrane Proteins / metabolism
  • Mitochondrial Proteins / metabolism
  • Zebrafish / genetics
  • Zebrafish / metabolism


  • Acetylcysteine
  • Cysteamine
  • Azides
  • Membrane Proteins
  • Mitochondrial Proteins
  • Electron Transport Complex IV
  • Glutathione
  • Lactates