title: Reduced activity and protein expression of NOS in R6/2 HD transgenic mice: effects of L-NAME on symptom progression.
abstract: Previous work found that dietary l-arginine alters symptom progression in mice transgenic for Huntington's disease (HD), and that cerebral blood flow (CBF) is abnormal in early stage HD patients. Both of these findings potentially implicate nitric oxide (NO) and its converting enzyme, nitric oxide synthase (NOS), in HD. The current experiment found that both NOS enzymatic activity and neuronal NOS (nNOS) protein expression were reduced (P<0.05) in R6/2 HD transgenic mice compared to non-HD controls (CON). Conversely, inducible NOS (iNOS) protein expression was not significantly different between groups. The changes in nNOS were accompanied by changes in protein expression of calmodulin kinase II (CaMKII) (P<0.05) and calmodulin kinase IV (CaMKIV) (P<0.05). Protein expression of 3-nitrotyrosine (3-NT), a marker for the neurotoxin peroxynitrite, was slightly increased in non-drug treated HD and was accompanied by increased immunostaining of 3-NT in cells adhering to the vasculature and choroid plexus. Mice that received the broad-spectrum NOS inhibitor N(g)-nitro-L-arginine methyl ester hydrochloride (L-NAME) via their drinking water had reduced NOS enzyme activity. NOS activity varied as a function of L-NAME dose, was virtually eliminated in the 500-mg/l groups, and correlated (P<0.05) with the behavioral scores as revealed by regression and correlation analyses. High dose L-NAME (500 mg/l) accelerated symptom onset in HD transgenics. These results support the hypothesis that nNOS activity and NO production are abnormal in HD, this in the setting of a more global dysregulation of calcium protein expression. Taken collectively with earlier data from our laboratory demonstrating abnormal CBF findings in early-stage HD patients, these results suggest that abnormalities in NOS function may significantly contribute to the neurodegeneration found in HD.
authors.affiliation: Department of Psychiatry, University of Connecticut Medical School, 263 Farmington Avenue, Farmington, CT 06030-2103, USA. deckel@psychiatry.uchc.edu
authors.forename: A W
authors.initials: AW
authors.lastname: Deckel
authors.forename: A
authors.initials: A
authors.lastname: Gordinier
authors.forename: D
authors.initials: D
authors.lastname: Nuttal
authors.forename: V
authors.initials: V
authors.lastname: Tang
authors.forename: C
authors.initials: C
authors.lastname: Kuwada
authors.forename: R
authors.initials: R
authors.lastname: Freitas
authors.forename: K A
authors.initials: KA
authors.lastname: Gary
mesh_terms: D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects; D001835:Body Weight / Q000187:drug effects; D001921:Brain / Q000187:drug effects / Q000201:enzymology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000096:biosynthesis; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D006816:Huntington Disease / Q000201:enzymology* / Q000235:genetics* / Q000503:physiopathology; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009043:Motor Activity / Q000187:drug effects; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D052248:Nitric Oxide Synthase Type I; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000096:biosynthesis
chemical_list: D004791:Enzyme Inhibitors; C002744:3-nitrotyrosine; D014443:Tyrosine; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; C496321:Nos1 protein, mouse; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D019331:NG-Nitroarginine Methyl Ester