Focal cerebral ischemia in the TNFalpha-transgenic rat

Background: To determine if chronic elevation of the inflammatory cytokine, tumor necrosis factor-alpha (TNF alpha), will affect infarct volume or cortical perfusion after focal cerebral ischemia. Methods: Transgenic (TNF alpha-Tg) rats overexpressing the murine TNF alpha gene in brain were prepared by injection of mouse DNA into rat oocytes. Brain levels of TNF alpha mRNA and protein were measured and compared between TNF alpha-Tg and non-transgenic (non-Tg) littermates. Mean infarct volume was calculated 24 hours or 7 days after one hour of reversible middle cerebral artery occlusion (MCAO). Cortical perfusion was monitored by laser-Doppler flowmetry (LDF) during MCAO. Cortical vascular density was quantified by stereology. Post-ischemic cell death was assessed by immunohistochemistry and regional measurement of caspase-3 activity or DNA fragmentation. Unpaired t tests or analysis of variance with post hoc tests were used for comparison of group means. Results: In TNF alpha-Tg rat brain, the aggregate mouse and rat TNF alpha mRNA level was fourfold higher than in non-Tg littermates and the corresponding TNF alpha protein level was increased fivefold (p <= 0.01). Infarct volume was greater in TNF alpha-Tg rats than in non-Tg controls at 24 hours (p <= 0.05) and 7 days (p <= 0.01). Within the first 10 minutes of MCAO, cortical perfusion measured by LDF was reduced in TNF alpha-Tg rats (p <= 0.05). However, regional vascular density was equivalent between TNF alpha-Tg and non-Tg animals (p = NS). Neural cellular apoptosis was increased in transgenic animals as shown by elevated caspase-3 activity (p = 0.05) and DNA fragmentation (p <= 0.001) at 24 hours. Conclusion: Chronic elevation of TNF alpha protein in brain increases susceptibility to ischemic injury but has no effect on vascular density. TNF alpha-Tg animals are more susceptible to apoptotic cell death after MCAO than are non-Tg animals. We conclude that the TNF alpha-Tg rat is a valuable new tool for the study of cytokine-mediated ischemic brain injury.