Focal cerebral ischemia in the TNFalpha-transgenic rat

L. Creed Pettigrew; Mark S. Kindy; Stephen Scheff; Joe E. Springer; Richard J. Kryscio; Yizhao Li; David S. Grass

doi:10.1186/1742-2094-5-47

Focal cerebral ischemia in the TNFalpha-transgenic rat

L. Creed Pettigrew, Mark S. Kindy, Stephen Scheff, Joe E. Springer, Richard J. Kryscio, Yizhao Li, David S. Grass

Research output: Contribution to journal › Article › peer-review

53 Scopus citations

Abstract

Background: To determine if chronic elevation of the inflammatory cytokine, tumor necrosis factor-α (TNFα), will affect infarct volume or cortical perfusion after focal cerebral ischemia. Methods: Transgenic (TNFα-Tg) rats overexpressing the murine TNFα gene in brain were prepared by injection of mouse DNA into rat oocytes. Brain levels of TNFα mRNA and protein were measured and compared between TNFα-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 TNFα-Tg rat brain, the aggregate mouse and rat TNFα mRNA level was fourfold higher than in non-Tg littermates and the corresponding TNFα protein level was increased fivefold (p ≤ 0.01). Infarct volume was greater in TNFα-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 TNFα-Tg rats (p ≤ 0.05). However, regional vascular density was equivalent between TNFα-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 elevationof TNFα protein in brain increases susceptibility to ischemic injury but has no effect on vascular density. TNFα-Tg animals are more susceptible to apoptotic cell death after MCAO than are non-Tg animals. We conclude that the TNFα-Tg rat is a valuable new tool for the study of cytokine-mediated ischemic brain injury.

Original language	English
Article number	47
Journal	Journal of Neuroinflammation
Volume	5
DOIs	https://doi.org/10.1186/1742-2094-5-47
State	Published - Oct 23 2008

Bibliographical note

Funding Information:
Financial support was provided by NIH/NINDS grants R01 NS047395 (LCP) and R01 NS039588 (MSK) and by a Visiting Research Scientist Award provided by Shandong University, China (YL) for training at the University of Kentucky in Lexington, Kentucky. Dr. George Kollias prepared the microinjection construct containing the murine TNFα gene and its promoter that was used to make the transgenic animal. Mr. Richard Coffee constructed the animal at Xenogen Biosciences in Cranbury, New Jersey. We thank Mary Louise Holtz, Ph.D., Xaio-Hong Song, M.D., Susan D. Crad-dock, and Ravikumar Rao for technical contributions to this work and Sherry Chandler Williams, ELS, for editing the manuscript and preparing the illustrations.

ASJC Scopus subject areas

Neuroscience (all)
Immunology
Neurology
Cellular and Molecular Neuroscience

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10.1186/1742-2094-5-47

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title = "Focal cerebral ischemia in the TNFalpha-transgenic rat",

abstract = "Background: To determine if chronic elevation of the inflammatory cytokine, tumor necrosis factor-α (TNFα), will affect infarct volume or cortical perfusion after focal cerebral ischemia. Methods: Transgenic (TNFα-Tg) rats overexpressing the murine TNFα gene in brain were prepared by injection of mouse DNA into rat oocytes. Brain levels of TNFα mRNA and protein were measured and compared between TNFα-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 TNFα-Tg rat brain, the aggregate mouse and rat TNFα mRNA level was fourfold higher than in non-Tg littermates and the corresponding TNFα protein level was increased fivefold (p ≤ 0.01). Infarct volume was greater in TNFα-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 TNFα-Tg rats (p ≤ 0.05). However, regional vascular density was equivalent between TNFα-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 elevationof TNFα protein in brain increases susceptibility to ischemic injury but has no effect on vascular density. TNFα-Tg animals are more susceptible to apoptotic cell death after MCAO than are non-Tg animals. We conclude that the TNFα-Tg rat is a valuable new tool for the study of cytokine-mediated ischemic brain injury.",

author = "Pettigrew, {L. Creed} and Kindy, {Mark S.} and Stephen Scheff and Springer, {Joe E.} and Kryscio, {Richard J.} and Yizhao Li and Grass, {David S.}",

note = "Funding Information: Financial support was provided by NIH/NINDS grants R01 NS047395 (LCP) and R01 NS039588 (MSK) and by a Visiting Research Scientist Award provided by Shandong University, China (YL) for training at the University of Kentucky in Lexington, Kentucky. Dr. George Kollias prepared the microinjection construct containing the murine TNFα gene and its promoter that was used to make the transgenic animal. Mr. Richard Coffee constructed the animal at Xenogen Biosciences in Cranbury, New Jersey. We thank Mary Louise Holtz, Ph.D., Xaio-Hong Song, M.D., Susan D. Crad-dock, and Ravikumar Rao for technical contributions to this work and Sherry Chandler Williams, ELS, for editing the manuscript and preparing the illustrations.",

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T1 - Focal cerebral ischemia in the TNFalpha-transgenic rat

AU - Pettigrew, L. Creed

AU - Kindy, Mark S.

AU - Scheff, Stephen

AU - Springer, Joe E.

AU - Kryscio, Richard J.

AU - Li, Yizhao

AU - Grass, David S.

N1 - Funding Information: Financial support was provided by NIH/NINDS grants R01 NS047395 (LCP) and R01 NS039588 (MSK) and by a Visiting Research Scientist Award provided by Shandong University, China (YL) for training at the University of Kentucky in Lexington, Kentucky. Dr. George Kollias prepared the microinjection construct containing the murine TNFα gene and its promoter that was used to make the transgenic animal. Mr. Richard Coffee constructed the animal at Xenogen Biosciences in Cranbury, New Jersey. We thank Mary Louise Holtz, Ph.D., Xaio-Hong Song, M.D., Susan D. Crad-dock, and Ravikumar Rao for technical contributions to this work and Sherry Chandler Williams, ELS, for editing the manuscript and preparing the illustrations.

PY - 2008/10/23

Y1 - 2008/10/23

N2 - Background: To determine if chronic elevation of the inflammatory cytokine, tumor necrosis factor-α (TNFα), will affect infarct volume or cortical perfusion after focal cerebral ischemia. Methods: Transgenic (TNFα-Tg) rats overexpressing the murine TNFα gene in brain were prepared by injection of mouse DNA into rat oocytes. Brain levels of TNFα mRNA and protein were measured and compared between TNFα-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 TNFα-Tg rat brain, the aggregate mouse and rat TNFα mRNA level was fourfold higher than in non-Tg littermates and the corresponding TNFα protein level was increased fivefold (p ≤ 0.01). Infarct volume was greater in TNFα-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 TNFα-Tg rats (p ≤ 0.05). However, regional vascular density was equivalent between TNFα-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 elevationof TNFα protein in brain increases susceptibility to ischemic injury but has no effect on vascular density. TNFα-Tg animals are more susceptible to apoptotic cell death after MCAO than are non-Tg animals. We conclude that the TNFα-Tg rat is a valuable new tool for the study of cytokine-mediated ischemic brain injury.

AB - Background: To determine if chronic elevation of the inflammatory cytokine, tumor necrosis factor-α (TNFα), will affect infarct volume or cortical perfusion after focal cerebral ischemia. Methods: Transgenic (TNFα-Tg) rats overexpressing the murine TNFα gene in brain were prepared by injection of mouse DNA into rat oocytes. Brain levels of TNFα mRNA and protein were measured and compared between TNFα-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 TNFα-Tg rat brain, the aggregate mouse and rat TNFα mRNA level was fourfold higher than in non-Tg littermates and the corresponding TNFα protein level was increased fivefold (p ≤ 0.01). Infarct volume was greater in TNFα-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 TNFα-Tg rats (p ≤ 0.05). However, regional vascular density was equivalent between TNFα-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 elevationof TNFα protein in brain increases susceptibility to ischemic injury but has no effect on vascular density. TNFα-Tg animals are more susceptible to apoptotic cell death after MCAO than are non-Tg animals. We conclude that the TNFα-Tg rat is a valuable new tool for the study of cytokine-mediated ischemic brain injury.

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Focal cerebral ischemia in the TNFalpha-transgenic rat

Abstract

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