Abstract
Glutaryl-CoA dehydrogenase deficiency (also known as glutaric aciduria type I) is an autosomal, recessively inherited neurometabolic disorder with a distinct neuropathology characterized by acute encephalopathy during a vulnerable period of brain development. Neuronal damage in this disease was demonstrated to involve N-methyl-d-aspartate (NMDA) receptor-mediated neurotoxicity of the endogenously accumulating metabolite 3-hydroxyglutarate (3-OH-GA). However, it remained unclear whether NMDA receptors are directly or indirectly activated and whether 3-OH-GA disturbs the intracellular Ca2+ homeostasis. Here we report that 3-OH-GA activated recombinant NMDA receptors (e.g. NR1/NR2A) but not recombinant α-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptors (e.g. GluR-A/GluR-B) in HEK293 cells. Fluorescence microscopy using fura-2 as Ca2+ indicator revealed that 3-OH-GA increased intracellular Ca2+ concentrations in the presence of extracellular Ca2+ in cultured chick neurons. Similar to glutamate-induced cell damage, 3-OH-GA neurotoxicity was modulated by extracellular Na+. The large cation N-methyl-d-glucamine, which does not permeate NMDA receptor channels, enhanced 3-OH-GA-induced Ca2+ increase and cell damage. In contrast, 3-OH-GA-induced neurotoxicity was reduced after replacement of Na+ by Li+, which permeates NMDA channels but does not affect the Na+/Ca2+ exchanger in the plasma membrane. Spectrophotometric analysis of respiratory chain complexes I–V in submitochondrial particles from bovine heart revealed only a weak inhibition of 3-OH-GA on complex V at the highest concentration tested (10 mM). In conclusion, the present study revealed that NMDA receptor activation and subsequent disturbance of Ca2+ homeostasis contribute to 3-OH-GA-induced cell damage.
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Abbreviations
- 3-OH-GA:
-
3-hydroxyglutarate
- AMPA:
-
α-amino-3-hydroxy-5-methyl-4-isoxazole propionate
- CNQX:
-
6-cyano-7-nitroquinoxaline-2,3-dione disodium salt
- D-AP5:
-
(D)-(−)-2-amino-5-phosphonopentanoic acid
- GCDH:
-
glutaryl-CoA dehydrogenase
- GDD:
-
glutaryl-CoA dehydrogenase deficiency
- HBS:
-
HEPES-buffered saline
- MK-801:
-
(5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine
- NaCaX:
-
Na+/Ca2+ exchanger of the plasma membrane
- NMDA:
-
N-methyl-d-aspartate
- NMG:
-
N-methyl-d-glucamine
- PM:
-
plasma membrane
- ROS:
-
reactive oxygen species
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Acknowledgements
The authors thank P.H. Seeburg (Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Heidelberg, Germany), H. Monyer (Department of Clinical Neurobiology, University of Heidelberg, Germany), and C.-M. Becker (Department of Biochemistry and Molecular Medicine, University of Erlangen, Germany) for fruitful discussions. We also thank S. Engel, S. Grünewald, and S. Exner-Camps for excellent technical support.
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Supported by a grant from the Deutsche Forschungsgemeinschaft to S.K. (KO 2010/1-1) and by a junior grant from the University of Heidelberg to F.H. (Nr. 12/2001).S.K. and G.K. contributed equally to the study.
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Kölker, S., Köhr, G., Ahlemeyer, B. et al. Ca2+ and Na+ Dependence of 3-Hydroxyglutarate-Induced Excitotoxicity in Primary Neuronal Cultures from Chick Embryo Telencephalons. Pediatr Res 52, 199–206 (2002). https://doi.org/10.1203/00006450-200208000-00011
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DOI: https://doi.org/10.1203/00006450-200208000-00011
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