title: Higher Vulnerability of Menadione-Exposed Cortical Astrocytes of Glutaryl-CoA Dehydrogenase Deficient Mice to Oxidative Stress, Mitochondrial Dysfunction, and Cell Death: Implications for the Neurodegeneration in Glutaric Aciduria Type I.
abstract: Patients affected by glutaric aciduria type I (GA-I) show progressive cortical leukoencephalopathy whose pathogenesis is poorly known. In the present work, we exposed cortical astrocytes of wild-type (Gcdh+/+) and glutaryl-CoA dehydrogenase knockout (Gcdh-/-) mice to the oxidative stress inducer menadione and measured mitochondrial bioenergetics, redox homeostasis, and cell viability. Mitochondrial function (MTT and JC1-mitochondrial membrane potential assays), redox homeostasis (DCFH oxidation, nitrate and nitrite production, GSH concentrations and activities of the antioxidant enzymes SOD and GPx), and cell death (propidium iodide incorporation) were evaluated in primary cortical astrocyte cultures of Gcdh+/+and Gcdh-/-mice unstimulated and stimulated by menadione. We also measured the pro-inflammatory response (TNFα levels, IL1-β and NF-ƙB) in unstimulated astrocytes obtained from these mice. Gcdh-/-mice astrocytes were more vulnerable to menadione-induced oxidative stress (decreased GSH concentrations and altered activities of the antioxidant enzymes), mitochondrial dysfunction (decrease of MTT reduction and JC1 values), and cell death as compared with Gcdh+/+astrocytes. A higher inflammatory response (TNFα, IL1-β and NF-ƙB) was also observed in Gcdh-/-mice astrocytes. These data indicate a higher susceptibility of Gcdh-/-cortical astrocytes to oxidative stress and mitochondrial dysfunction, probably leading to cell death. It is presumed that these pathomechanisms may contribute to the cortical leukodystrophy observed in GA-I patients.
authors: Departamento e PPG Bioquímica, ICBS/Universidade Federal do Rio Grande do Sul (UFRGS), Rua Ramiro Barcelos N° 2600, Anexo, Porto Alegre, RS, CEP90035-003, Brazil.
authors: Marília Danyelle Nunes
authors: MDN
authors: Rodrigues
authors: Departamento e PPG Bioquímica, ICBS/Universidade Federal do Rio Grande do Sul (UFRGS), Rua Ramiro Barcelos N° 2600, Anexo, Porto Alegre, RS, CEP90035-003, Brazil.
authors: Bianca
authors: B
authors: Seminotti
authors: Departamento e PPG Bioquímica, ICBS/Universidade Federal do Rio Grande do Sul (UFRGS), Rua Ramiro Barcelos N° 2600, Anexo, Porto Alegre, RS, CEP90035-003, Brazil.
authors: Ângela
authors: Â
authors: Zanatta
authors: Departamento e PPG Bioquímica, ICBS/Universidade Federal do Rio Grande do Sul (UFRGS), Rua Ramiro Barcelos N° 2600, Anexo, Porto Alegre, RS, CEP90035-003, Brazil.
authors: Aline
authors: A
authors: de Mello Gonçalves
authors: Departamento e PPG Bioquímica, ICBS/Universidade Federal do Rio Grande do Sul (UFRGS), Rua Ramiro Barcelos N° 2600, Anexo, Porto Alegre, RS, CEP90035-003, Brazil.
authors: Bruna
authors: B
authors: Bellaver
authors: Departamento e PPG Bioquímica, ICBS/Universidade Federal do Rio Grande do Sul (UFRGS), Rua Ramiro Barcelos N° 2600, Anexo, Porto Alegre, RS, CEP90035-003, Brazil.
authors: Alexandre Umpierrez
authors: AU
authors: Amaral
authors: Departamento e PPG Bioquímica, ICBS/Universidade Federal do Rio Grande do Sul (UFRGS), Rua Ramiro Barcelos N° 2600, Anexo, Porto Alegre, RS, CEP90035-003, Brazil.
authors: André
authors: A
authors: Quincozes-Santos
authors: Department of Pediatrics, University of Colorado Denver, Aurora, CO, USA.
authors: Stephen Irwin
authors: SI
authors: Goodman
authors: Department of Pediatrics, University of Colorado Denver, Aurora, CO, USA.
authors: Michael
authors: M
authors: Woontner
authors: Departamento e PPG Bioquímica, ICBS/Universidade Federal do Rio Grande do Sul (UFRGS), Rua Ramiro Barcelos N° 2600, Anexo, Porto Alegre, RS, CEP90035-003, Brazil.
authors: Diogo Onofre
authors: DO
authors: Souza
authors: Departamento e PPG Bioquímica, ICBS/Universidade Federal do Rio Grande do Sul (UFRGS), Rua Ramiro Barcelos N° 2600, Anexo, Porto Alegre, RS, CEP90035-003, Brazil. mwajner@ufrgs.br.|Serviço de Genética Médica, Hospital de Clínicas de Porto Alegre, Porto Alegre, RS, Brazil. mwajner@ufrgs.br.
authors: Moacir
authors: M
authors: Wajner
mesh_terms: D000592:Amino Acid Metabolism, Inborn Errors / Q000201:enzymology / Q000473:pathology*; D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D001253:Astrocytes / Q000187:drug effects / Q000473:pathology*; D001928:Brain Diseases, Metabolic / Q000201:enzymology / Q000473:pathology*; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002540:Cerebral Cortex / Q000473:pathology*; D005452:Fluoresceins / Q000378:metabolism; D050770:Glutaryl-CoA Dehydrogenase / Q000172:deficiency*; D005979:Glutathione Peroxidase / Q000378:metabolism; D018836:Inflammation Mediators / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D009410:Nerve Degeneration / Q000201:enzymology / Q000473:pathology*; D009569:Nitric Oxide / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D024483:Vitamin K 3 / Q000633:toxicity*
chemical_list: D000975:Antioxidants; D005452:Fluoresceins; D018836:Inflammation Mediators; D017382:Reactive Oxygen Species; C065013:2',7'-dichlorodihydrofluorescein; D009569:Nitric Oxide; D024483:Vitamin K 3; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D050770:Glutaryl-CoA Dehydrogenase
keywords: Gcdh −/− mice astrocytes; Glutaric aciduria type I; Mitochondrial dysfunction; Oxidative stress; Pro-inflammatory response