Nitric oxide synthase 3-mediated neurodegeneration after intracerebral gene delivery.
In Alzheimer disease (AD), increased nitric oxide synthase 3 (NOS3) expression correlates with apoptosis in cortical neurons and colocalizes with amyloid precursor protein (APP)-amyloid beta (Abeta) deposits in the brain. In the present study we examined the potential role of NOS3 in relation to AD-type neurodegeneration using an in vivo model of gene delivery. Long Evans rat pups were given a single intracerebral injection of recombinant plasmid DNA containing the human NOS3 cDNA (p-hNOS3) or the luciferase (p-Luc) gene as a negative control, and complexed with polyamine reagent. Overexpression of NOS3 in the brain increased the levels of APP, APP-Abeta, p53, Tau, glial fibrillary acidic protein, and peroxisome proliferator activated receptors (PPAR) delta and gamma and decreased the levels of Hu (neuronal marker) mRNA, phosphorylated glycogen synthase kinase 3beta, ATP synthase, and choline acetyltransferase expression as demonstrated by real-time quantitative reverse-transcribed polymerase chain reaction, Western blot analysis, or immunohistochemical staining. These effects of NOS3 overexpression were accompanied by increased single-stranded DNA immunoreactivity, reflecting DNA damage. The results suggest that increased cerebral expression of NOS3 causes several molecular abnormalities related to AD-type neurodegeneration, including oxidative stress, mitochondrial dysfunction, and impaired acetylcholine homeostasis. The coexisting increases in PPAR-delta and -gamma expression suggest that the adverse effects of NOS3 overexpression may be abated by PPAR agonist treatment.
de la Monte
Suzanne M
SM
Department of Medicine, Rhode Island Hospital, Brown Medical School, Providence, Rhode Island 02903, USA. Suzanne_DeLaMonte_MD@Brown.edu
Jhaveri
Ami
A
Maron
Bradley A
BA
Wands
Jack R
JR
D016564:Amyloid beta-Protein Precursor / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western / Q000379:methods; D001921:Brain / Q000378:metabolism; D004195:Disease Models, Animal; D015870:Gene Expression / Q000502:physiology; D005786:Gene Expression Regulation / Q000235:genetics; D018014:Gene Transfer Techniques; D006801:Humans; D009419:Nerve Tissue Proteins / Q000378:metabolism; D019636:Neurodegenerative Diseases / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D052250:Nitric Oxide Synthase Type III / Q000008:administration & dosage* / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D020318:Rats, Long-Evans; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013997:Time Factors; D016875:tau Proteins / Q000378:metabolism
D016564:Amyloid beta-Protein Precursor; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D016875:tau Proteins; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III