title: A novel, rapid, computerized method for quantitation of neuronal damage in a rat model of stroke.
abstract: Determination of extent of infarction in animal models of cerebral ischemia is most commonly achieved by either classical histology (thionin staining) and light microscopy or staining with 2,3, 5-triphenyltetrazolium chloride (TTC). These techniques have limitations and we now describe a novel technique and its validation for assessment of the neuroprotective activity of AM-36, a novel arylalkypiperazine compound with combined antioxidant and sodium channel blocking activity. AM-36 (1.8 mg/kg i.p.) or vehicle, was administered 30 min, 24 and 48 h after endothelin-1-induced middle cerebral artery occlusion in conscious rats. Rats were killed at 72 h, brains removed and frozen in liquid nitrogen prior to coronal sectioning. Using a simple apparatus relying on basic principles of light propagation and a computerised image analysis system, ischemic damage in unstained slide-mounted sections was clearly visualised and measured. AM-36 significantly reduced the area of infarct in both cortex and striatum. The method was verified by thionin staining, and light microscopy. Linear regression analysis showed a highly significant correlation between methods at 72 h for infarct area in the cortex and striatum. Highly significant correlations between methods were found at 3 and 24 h after ischemia. Our method quickly and clearly delineates areas of damage in a manner superior to conventional staining methods.
authors.affiliation: Department of Pharmacology, Monash University, PO Box 13E, Clayton 3800, Australia. jennifer.callaway@med.monash.edu.au
authors.forename: J K
authors.initials: JK
authors.lastname: Callaway
authors.forename: M J
authors.initials: MJ
authors.lastname: Knight
authors.forename: D J
authors.initials: DJ
authors.lastname: Watkins
authors.forename: P M
authors.initials: PM
authors.lastname: Beart
authors.forename: B
authors.initials: B
authors.lastname: Jarrott
authors.forename: P M
authors.initials: PM
authors.lastname: Delaney
mesh_terms: D000818:Animals; D002545:Brain Ischemia / Q000188:drug therapy / Q000473:pathology* / Q000503:physiopathology; D002540:Cerebral Cortex / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology; D004195:Disease Models, Animal; D007091:Image Processing, Computer-Assisted; D008297:Male; D018715:Microscopy, Video / Q000295:instrumentation* / Q000379:methods; D017072:Neostriatum / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology; D009410:Nerve Degeneration / Q000188:drug therapy / Q000473:pathology* / Q000503:physiopathology; D018696:Neuroprotective Agents / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D020521:Stroke / Q000188:drug therapy / Q000473:pathology* / Q000503:physiopathology; D013997:Time Factors
chemical_list: D018696:Neuroprotective Agents; D010879:Piperazines; C401121:AM 36