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Volume 16, Number 21, Issue of November 1, 1996 pp. 6795-6806
Copyright ©1996 Society for Neuroscience

Damage, Repair, and Mutagenesis in Nuclear Genes after Mouse Forebrain Ischemia-Reperfusion

Received June 20, 1996; revised Aug. 2, 1996; accepted Aug. 13, 1996.

Philip K. Liu1, Chung Y. Hsu2, Miral Dizdaroglu3, Robert A. Floyd4, Yoke W. Kow5, Asuman Karakaya3, 6, Lois E. Rabow5, and Jian-K. Cui1

1 Laboratory of Neurobiology, Division of Restorative Neurology and Human Neurobiology, Baylor College of Medicine, Houston, Texas 77030, 2 Department of Neurology, Washington University, St. Louis, Missouri 63110, 3 Chemical Science and Technology Laboratory, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, 4 Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104-5046, 5 Department of Radiation Oncology, Emory University School of Medicine, Atlanta, Georgia 30335, and 6 Faculty of Pharmacy, University of Ankara, Ankara, Turkey

To determine whether oxidative stress after cerebral ischemia-reperfusion affects genetic stability in the brain, we studied mutagenesis after forebrain ischemia-reperfusion in Big Blue transgenic mice (male C57BL/6 strain) containing a reporter lacI gene, which allows detection of mutation frequency. The frequency of mutation in this reporter lacI gene increased from 1.5 to 7.7 (per 100,000) in cortical DNA after 30 min of forebrain ischemia and 8 hr of reperfusion and remained elevated at 24 hr reperfusion. Eight DNA lesions that are characteristic of DNA damage mediated by free radicals were detected. Four mutagenic lesions (2,6-diamino-4-hydroxy-5-formamidopyrimidine, 8-hydroxyadenine, 5-hydroxycytosine, and 8-hydroxyguanine) examined by gas chromatography/mass spectrometry and one corresponding 8-hydroxy-2'-deoxyguanosine by a method of HPLC with electrochemical detection increased in cortical DNA two- to fourfold (p < 0.05) during 10-20 min of reperfusion. The damage to gamma -actin and DNA polymerase-beta genes was detected within 20 min of reperfusion based on the presence of formamidopyrimidine DNA N-glycosylase-sensitive sites. These genes became resistant to the glycosylase within 4-6 hr of reperfusion, suggesting a reduction in DNA damage and presence of DNA repair in nuclear genes. These results suggest that nuclear genes could be targets of free radicals.

Key words: apoptosis; brain; mutation; dementia; oxidative stress; stroke




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