Inducible nitric oxide synthase does not mediate brain damage after transient focal cerebral ischemia in mice

• Published in: Journal of cerebral blood flow and metabolism 2008-03, Vol.28 (3), p.526-539
• Main Authors: Prüss, Harald, Prass, Konstantin, Ghaeni, Leyli, Milosevic, Milan, Muselmann, Claudia, Freyer, Dorette, Royl, Georg, Reuter, Uwe, Baeva, Nevena, Dirnagl, Ulrich, Meisel, Andreas, Priller, Josef
• Format: Article
• Language: English
• Subjects: Alternative Splicing; Animals; Biological and medical sciences; Brain Chemistry - genetics; Brain Ischemia - enzymology; Brain Ischemia - etiology; Brain Ischemia - metabolism; Exons; Ischemic Attack, Transient - enzymology; Ischemic Attack, Transient - etiology; Ischemic Attack, Transient - metabolism; Lung - enzymology; Medical sciences; Mice; Mice, Knockout; Neurology; Neuropharmacology; Neuroprotective agent; Nitric Oxide Synthase Type II - deficiency; Nitric Oxide Synthase Type II - genetics; Pharmacology. Drug treatments; RNA, Messenger - genetics; Up-Regulation - genetics; Vascular diseases and vascular malformations of the nervous system
• ISSN: 0271-678X; 1559-7016
• DOI: 10.1038/sj.jcbfm.9600550

Nitric oxide produced by the inducible nitric oxide synthase (iNOS) is believed to participate in the pathogenic events after cerebral ischemia. In this study, we examined the expression of iNOS in the brain after transient focal cerebral ischemia in mice. We detected differential expression of exons 2 and 3 of iNOS mRNA (16-fold upregulation at 24 to 72 h after middle cerebral artery occlusion, MCAO) compared with exons 6 to 8, 12 to 14, 21 to 22, and 26 to 27 (2- to 5-fold upregulation after 72 and 96 h), which would be compatible with alternative splicing. Expression levels of iNOS mRNA were too low for detection by the Northern blot analysis. Using specific antibodies, we did not detect any iNOS immunoreactivity in the mouse brain 1 to 5 days after MCAO, although we detected iNOS immunoreactivity in the lungs of mice with stroke-associated pneumonia, and in mouse and rat dura mater after lipopolysaccharide administration. In chimeric iNOS-deficient mice transplanted with wild-type bone marrow (BM) cells expressing the green fluorescent protein (GFP) or in wild-type mice transplanted with GFP+ iNOS-deficient BM cells, no expression of iNOS was detected in GFP+ leukocytes invading the ischemic brain or in resident brain cells. Moreover, both experimental groups did not show any differences in infarct size. Analysis of three different strains of iNOS-deficient mice and wild-type controls confirmed that infarct size was independent of iNOS deletion, but strongly confounded by the genetic background of mouse strains. In conclusion, our data suggest that iNOS is not a universal mediator of brain damage after cerebral ischemia.