Microvascular basal lamina injury after experimental focal cerebral ischemia and reperfusion in the rat

To define the location and extent of microvascular damage of the basal lamina after cerebral ischemia and reperfusion in rats. the authors subjected animals (n = 16) to 3 hours of focal cerebral ischemia and 24 hours of reperfusion using the suture middle cerebral artery occlusion model: sham-operated animals served as controls (n = 6). The Western blot technique was used to define the collagen type IV protein content in various brain regions, whereas immunohistochemistry identified microvascular basal lamina loss (anticollagen type IV staining). The extent of damage was quantified by automatic morphometric video-imaging analysis. Statistical analysis was based on the Mann-Whitney test and the paired Student's t-test. The ischemic hemisphere showed a reduction of the collagen type IV protein content after ischemia and reperfusion in the Western blot (reduction compared with the nonischemic side: total hemisphere. 33% +/- 6%: basal ganglia, 49% +/- 4% cortex, 25% +/- 7%: P < 0.01). There was also a decrease in the number of cerebral microvessels between the ischemic and nonis-chemic hemispheres (16% +/- 3%), cortical (14% +/- 3%), and basal ganglia areas 21% +/- 4%) (P < 0.01). Besides a reduction of the vessel number. there was also a loss in basal lamina antigen-positive stained area in ischemic areas (hemisphere, 20% +/- 2% : cortex, 8% +/- 3%: basal ganglia, 31% +/- 3% P < 0.001 Cortical areas had a less pronounced basal lamina loss than basal ganglia (P < 0.05). For the first time, microvascular basal lamina damage, indicated by collagen type IV loss, is proven in rats by biochemical and morphometric analysis. These changes are comparable with those found in nonhuman primates. The authors report novel data regarding microvascular ischemic changes in the cortex. These data provide a basis for future experiment,., to determine the mechanisms of ischemic microvascular damage and to devise new therapeutic strategies.