title: Pathological implications of iNOS expression in central white matter: an ex vivo study of optic nerves from rats with experimental allergic encephalomyelitis.
abstract: Excessive nitric oxide (NO) production from the inducible isoform of nitric oxide synthase (iNOS) has been invoked as a causative factor in many neurodegenerative disorders, including multiple sclerosis. This hypothesis has been supported by in vitro studies showing that glial iNOS expression results in toxic NO concentrations (near 1 microm). To investigate the relevance of such findings, experiments were carried out ex vivo on optic nerves from rats with exacerbated experimental allergic encephalomyelitis, a model of multiple sclerosis. The nerves displayed characteristic immunopathology and expression of iNOS in macrophages and/or microglia and there was overt axonal damage in localized regions of the optic chiasm. The resulting NO levels in the optic nerve were sufficient to cause activation of guanylyl cyclase-coupled NO receptors, resulting in marked cGMP accumulation in axons throughout the nerve. Nevertheless, calibration of cGMP levels against those evoked by exogenous NO indicated that the nerves were not compromised metabolically and that their ambient NO concentration was only approximately 1 nm. Consistent with this observation, electrophysiological tests indicated that there was no ongoing malfunctioning of the type that can be elicited by high exogenous NO concentrations. It is concluded that, with iNOS expressed in physiological locations and levels, the tissue levels of NO remain at concentrations far lower than those shown to have toxic effects, despite continuous NO synthesis. The fact that NO can rise to much higher levels in dispersed cultures in vitro may be attributable to a deficiency in NO inactivation in such preparations.
authors.affiliation: The Wolfson Institute for Biomedical Research, University College London, Gower, Street, London WC1E 6BT, UK. g.garthwaite@ucl.ac.uk
authors.forename: G
authors.initials: G
authors.lastname: Garthwaite
authors.forename: A M
authors.initials: AM
authors.lastname: Batchelor
authors.forename: D A
authors.initials: DA
authors.lastname: Goodwin
authors.forename: A K
authors.initials: AK
authors.lastname: Hewson
authors.forename: K
authors.initials: K
authors.lastname: Leeming
authors.forename: Z
authors.initials: Z
authors.lastname: Ahmed
authors.forename: M L
authors.initials: ML
authors.lastname: Cuzner
authors.forename: J
authors.initials: J
authors.lastname: Garthwaite
mesh_terms: D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D001120:Arginine / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism; D039481:CD11b Antigen / Q000378:metabolism; D018801:CD2 Antigens / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D003671:DEET / Q000494:pharmacology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004558:Electric Stimulation / Q000379:methods; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000139:chemically induced / Q000201:enzymology* / Q000473:pathology*; D005260:Female; D006162:Guanylate Cyclase / Q000378:metabolism; D006168:Guinea Pigs; D006834:Hydrazines / Q000494:pharmacology; D007150:Immunohistochemistry / Q000379:methods; D008264:Macrophages / Q000201:enzymology* / Q000473:pathology; D046529:Microscopy, Electron, Transmission / Q000379:methods; D004676:Myelin Basic Protein / Q000378:metabolism; D009569:Nitric Oxide / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D009897:Optic Chiasm / Q000473:pathology / Q000648:ultrastructure; D009900:Optic Nerve / Q000187:drug effects / Q000201:enzymology / Q000473:pathology* / Q000648:ultrastructure; D009952:Ornithine / Q000031:analogs & derivatives* / Q000494:pharmacology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D051381:Rats; D011917:Rats, Inbred Lew; D013997:Time Factors
chemical_list: D015415:Biomarkers; D039481:CD11b Antigen; D018801:CD2 Antigens; D006834:Hydrazines; D004676:Myelin Basic Protein; C112653:PAPA NONOate; D010726:Phosphodiesterase Inhibitors; D003671:DEET; D009569:Nitric Oxide; C065027:N(G)-iminoethylornithine; D001120:Arginine; D009952:Ornithine; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D006162:Guanylate Cyclase; D006152:Cyclic GMP; D015056:1-Methyl-3-isobutylxanthine