Neurotoxicity Research

, Volume 33, Issue 3, pp 681–692 | Cite as

Experimental Evidence that In Vivo Intracerebral Administration of L-2-Hydroxyglutaric Acid to Neonatal Rats Provokes Disruption of Redox Status and Histopathological Abnormalities in the Brain

  • Rafael Teixeira Ribeiro
  • Ângela Zanatta
  • Alexandre Umpierrez Amaral
  • Guilhian Leipnitz
  • Francine Hehn de Oliveira
  • Bianca Seminotti
  • Moacir Wajner


Tissue accumulation of L-2-hydroxyglutaric acid (L-2-HG) is the biochemical hallmark of L-2-hydroxyglutaric aciduria (L-2-HGA), a rare neurometabolic inherited disease characterized by neurological symptoms and brain white matter abnormalities whose pathogenesis is not yet well established. L-2-HG was intracerebrally administered to rat pups at postnatal day 1 (P1) to induce a rise of L-2-HG levels in the central nervous system (CNS). Thereafter, we investigated whether L-2-HG in vivo administration could disturb redox homeostasis and induce brain histopathological alterations in the cerebral cortex and striatum of neonatal rats. L-2-HG markedly induced the generation of reactive oxygen species (increase of 2′,7′-dichloroflurescein-DCFH-oxidation), lipid peroxidation (increase of malondialdehyde concentrations), and protein oxidation (increase of carbonyl formation and decrease of sulfhydryl content), besides decreasing the antioxidant defenses (reduced glutathione-GSH) and sulfhydryl content in the cerebral cortex. Alterations of the activities of various antioxidant enzymes were also observed in the cerebral cortex and striatum following L-2-HG administration. Furthermore, L-2-HG-induced lipid peroxidation and GSH decrease in the cerebral cortex were prevented by the antioxidant melatonin and by the classical antagonist of NMDA glutamate receptor MK-801, suggesting the involvement of reactive species and of overstimulation of NMDA receptor in these effects. Finally, L-2-HG provoked significant vacuolation and edema particularly in the cerebral cortex with less intense alterations in the striatum that were possibly associated with the unbalanced redox homeostasis caused by this metabolite. Taken together, it is presumed that these pathomechanisms may underlie the neurological symptoms and brain abnormalities observed in the affected patients.


L-2-hydroxyglutaric aciduria L-2-hydroxyglutaric acid Redox homeostasis Cerebral cortex Striatum Histopathology 


Funding Information

This work was supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico #404883/2013-3, Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul #2266-2551/14-2, Pró-Reitoria de Pesquisa/Universidade Federal do Rio Grande do Sul #PIBIC 27613, FIPE/HCPA, and Financiadora de Estudos e Projetos/Rede Instituto Brasileiro de Neurociência #01.06.0842-00.

Compliance with Ethical Standards

Ethical Approval

The experimental protocol was approved by the Ethics Committee for Animal Research of the UFRGS, Porto Alegre, Brazil, and followed the “National Institutes of Health Guide for the Care and Use of Laboratory Animals” (NIH Publications No. 85–23, revised 2011).

Conflict of Interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Rafael Teixeira Ribeiro
    • 3
  • Ângela Zanatta
    • 1
  • Alexandre Umpierrez Amaral
    • 1
    • 2
  • Guilhian Leipnitz
    • 1
    • 3
  • Francine Hehn de Oliveira
    • 5
  • Bianca Seminotti
    • 1
  • Moacir Wajner
    • 1
    • 3
    • 4
  1. 1.Programa de Pós-Graduação em Ciências Biológicas: Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do SulPorto AlegreBrazil
  2. 2.Departamento de Ciências BiológicasUniversidade Regional Integrada do Alto Uruguai e das MissõesErechimBrazil
  3. 3.Departamento de Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do SulPorto AlegreBrazil
  4. 4.Serviço de Genética MédicaHospital de Clínicas de Porto AlegrePorto AlegreBrazil
  5. 5.Serviço de PatologiaHospital de Clinicas de Porto AlegrePorto AlegreBrazil

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