title: Evaluation of 3-nitrotyrosine as a marker for 3-nitropropionic acid-induced oxidative stress in Lewis and Wistar rats and strain-specific whole brain spheroid cultures.
abstract: The present study investigated whether 3-nitrotyrosine is an early marker for neurodegenerative processes involving oxidative stress. We characterized the 3-nitrotyrosine formation after 3-nitropropionic acid (3-NP) exposure in the whole brain spheroid culture model and in a rat model, using Lewis and Wistar rats. Increased 3-nitrotyrosine concentration in spheroid cultures from Lewis rats was observed at lower dose of and shorter exposure time to 3-NP as compared to alterations in glial fibrillary acidic protein concentration, decrease in glutamine synthetase activity or cell loss. Five days of exposure to 3-NP (5 mM) resulted in decreased staining of GABAergic processes, while neuronal nitric oxide synthase staining was preserved. In addition, staining of EAAC1, anti-2',3'-cyclic nucleotide 3'-phosphohydrolase and ED1 was diminished after treatment of spheroid cultures with 3-nitropropionic acid (5 mM), while isolectin B4 staining was increased. Dithiothreitol and vitamin E inhibited the increased formation of 3-nitrotyrosine. Interestingly, N(G)-nitro-L-arginine methyl ester increased the 3-nitrotyrosine formation. No increased 3-nitrotyrosine concentration was shown after exposure to 3-nitropropionic acid during 5 days in spheroid cultures obtained from Wistar rats. In the striatum of 3-NP-exposed Lewis and Wistar rats, no change in 3-nitrotyrosine concentration was observed, whereas only in Wistar rats the glial fibrillary acidic protein concentration was increased in addition to activation of microglial cells. It is concluded that 3-nitrotyrosine was a more sensitive marker for oxidative stress-induced neurodegeneration than glial fibrillary acidic protein and glutamine synthase in spheroid cell cultures of Lewis rats. Finally, the similarities between the 3-NP spheroid model and the vivo model indicate that the spheroid cultures provide a good alternative for chronic exposure of animals to neurotoxins.
authors.affiliation: European Graduate School of Neuroscience (Euron), Universiteit Maastricht, Department of Psychiatry and Neuropsychology, P.O. Box 616, 6200 MD, Maastricht, The Netherlands. c.teunissen@np.unimaas.nl
authors.forename: Charlotte E
authors.initials: CE
authors.lastname: Teunissen
authors.forename: Marjanne
authors.initials: M
authors.lastname: Markerink-van Ittersum
authors.forename: Chris
authors.initials: C
authors.lastname: de Bruijn
authors.forename: Harry W M
authors.initials: HW
authors.lastname: Steinbusch
authors.forename: Jan
authors.initials: J
authors.lastname: de Vente
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001522:Behavior, Animal / Q000187:drug effects; D015415:Biomarkers; D001923:Brain Chemistry / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005455:Fluorescent Antibody Technique; D005609:Free Radicals / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009498:Neurotoxins / Q000008:administration & dosage / Q000494:pharmacology*; D009574:Nitro Compounds; D018384:Oxidative Stress / Q000187:drug effects*; D011247:Pregnancy; D011422:Propionates / Q000008:administration & dosage / Q000494:pharmacology*; D051381:Rats; D011917:Rats, Inbred Lew; D017208:Rats, Wistar; D013045:Species Specificity; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D005609:Free Radicals; D005904:Glial Fibrillary Acidic Protein; D009498:Neurotoxins; D009574:Nitro Compounds; D011422:Propionates; C002744:3-nitrotyrosine; D014443:Tyrosine; C015392:3-nitropropionic acid; D019331:NG-Nitroarginine Methyl Ester