title: Assessment of induction of biliverdin reductase in a mouse model of middle cerebral artery occlusion.
abstract: Reproducible animal models of stroke are indispensable for investigation of pathogenesis and treatment of ischemic brain injury. Defined location and size of infarction as well as consistent production of neurological deficits make it possible to evaluate therapeutic potential of neuroprotective agents as well as to assess the impact of gene deletion [Nat. Med. 3 (1997) 1089; Science 265 (1994) 1883; Nat. Med. 4 (1998) 228] or overexpression [J. Neurochem. 72 (1999) 1187; J. Neurosci. 17 (1997) 7655] on neuroprotection in genetically altered mice. Ischemic stroke in mice can be reliably replicated by means of an open craniectomy exposure followed by permanent occlusion of the trunk and branches of the middle cerebral artery (MCA). Open craniectomy model is known to be statistically robust, yielding a coefficient of variation of <10%, and requiring minimal number of animals to validate the concept of statistical power. In the past, this model as well as some of its variants had been used in pivotal scientific studies to demonstrate impact of therapeutic genes on the course of ischemic neuronal injury [Neuron 13 (1994) 1017], as well as identification of 'culprit genes' responsible for progression of ischemic injury [J. Cerebr. Blood Flow Metab. 14 (1994) 887; Science 265 (1994) 1883; J. Neurosci. 17 (1997) 7655] through continuous recruitment of marginally ischemic penumbra into ischemic core [Trends Neurosci. 22 (1999) 391]. This protocol describes mapping of the ischemic penumbra using NADPH diaphorase staining as well as assessment of penumbral endogenous antioxidant reserves by detection of cellular biliverdin reductase mRNA and protein levels using immunocytochemistry and in situ hybridization histological techniques.
authors.affiliation: Department of Biochemistry and Biophysics, Box 712, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, NY 14642, USA.
authors.forename: N
authors.initials: N
authors.lastname: Panahian
authors.forename: M D
authors.initials: MD
authors.lastname: Maines
mesh_terms: D000818:Animals; D001157:Arterial Occlusive Diseases / Q000201:enzymology*; D002536:Cerebral Arteries; D004790:Enzyme Induction; D005260:Female; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D051379:Mice; D009252:NADPH Dehydrogenase / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D044925:Oxidoreductases Acting on CH-CH Group Donors; D012333:RNA, Messenger / Q000378:metabolism; D013194:Staining and Labeling
chemical_list: D012333:RNA, Messenger; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; C019789:biliverdin reductase; D009252:NADPH Dehydrogenase