Abstract
Glutaryl-CoA dehydrogenase deficiency is an autosomal recessively inherited neurometabolic disorder with a distinct neuropathology characterized by acute encephalopathic crises during a vulnerable period of brain development. 3-Hydroxyglutarate (3-OH-GA), which accumulates in affected patients, has been identified as an endogenous neurotoxin mediating excitotoxicity via N-methyl-d-aspartate receptors. As increased generation of reactive oxygen species (ROS) and nitric oxide (NO) plays an important role in excitotoxic neuronal damage, we investigated whether ROS and NO contribute to 3-OH-GA neurotoxicity. 3-OH-GA increased mitochondrial ROS generation in primary neuronal cultures from chick embryo telencephalons, which could be prevented by MK-801, confirming the central role of N-methyl-d-aspartate receptor stimulation in 3-OH-GA toxicity. ROS increase was reduced by α-tocopherol and—less effectively—by melatonin. α-Tocopherol revealed a wider time frame for neuroprotection than melatonin. Creatine also reduced neuronal damage and ROS formation but only if it was administered ≥6 h before 3-OH-GA. NO production revealed only a slight increase after 3-OH-GA incubation. NO synthase inhibitor Nω-nitro-l-arginine prevented NO increase but did not protect neurons against 3-OH-GA. The NO donor S-nitroso-N-acetylpenicillamine revealed no effect on 3-OH-GA toxicity at low concentrations (0.5–5 μM), whereas it potentiated neuronal damage at high concentrations (50–500 μM), suggesting that weak endogenous NO production elicited by 3-OH-GA did not affect neuronal viability. We conclude from our results that ROS generation contributes to 3-OH-GA neurotoxicity in vitro and that radical scavenging and stabilization of brain energy metabolism by creatine are hopeful new strategies in glutaryl-CoA dehydrogenase deficiency.
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Abbreviations
- 3-OH-GA:
-
3-hydroxyglutarate
- DHR:
-
dihydrorhodamine-123
- GCDH:
-
glutaryl-CoA dehydrogenase (EC 1.3.99.7)
- GDD:
-
glutaryl-CoA dehydrogenase deficiency
- HBS:
-
magnesium- and zinc-free HEPES buffered saline
- l-NAME:
-
Nω-nitro-l-arginine
- MK-801:
-
(5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a d]cyclohepten-5 10-imine
- NMDA:
-
N-methyl-d-aspartate
- NO:
-
nitric oxide
- nNOS:
-
neuronal nitric oxide synthase; synonym nitric oxide synthase type I (EC 1.14.13.39)
- NO−:
-
nitroxyl anion
- NO+:
-
nitrosonium ion
- ROS:
-
reactive oxygen species
- SNAP:
-
S-nitroso-n-acetylpenicillamine
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Supported by Deutsche Forschungsgemeinschaft (grant KO 2010/1-1 to S.K.).
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Kölker, S., Ahlemeyer, B., Krieglstein, J. et al. Contribution of Reactive Oxygen Species to 3-Hydroxyglutarate Neurotoxicity in Primary Neuronal Cultures from Chick Embryo Telencephalons. Pediatr Res 50, 76–82 (2001). https://doi.org/10.1203/00006450-200107000-00015
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DOI: https://doi.org/10.1203/00006450-200107000-00015
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