A novel, rapid, computerised method for quantitation of neuronal damage ina rat model of stroke

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 techni que and its validation for assessment of the neuroprotective activity of AM -36, a novel arylalkypiperazine compound with combined antioxidant and sodi um channel blocking activity. AM-36 (1.8 mg/kg i.p.) or vehicle, was admini stered 30 min, 24 and 48 h after endothelin-1-induced middle cerebral arter y occlusion in conscious rats. Rats were killed at 72 h, brains removed and frozen in liquid nitrogen prior to coronal sectioning. Using a simple appa ratus relying on basic principles of light propagation and a computerised i mage analysis system, ischemic damage in unstained slide-mounted sections w as clearly visualised and measured. AM-36 significantly reduced the area of infarct in both cortex and striatum. The method was verified by thionin st aining, and light microscopy. Linear regression analysis showed a highly si gnificant correlation between methods at 72 h for infarct area in the corte x and striatum. Highly significant correlations between methods were found at 3 and 24 h after ischemia. Our method quickly and clearly delineates are as of damage in a manner superior to conventional staining methods. (C) 200 0 Elsevier Science B.V. All rights reserved.