Hyperglycemia enhances DNA fragmentation after transient cerebral ischemia

Journal of Cerebral Blood Flow and Metabolism : Official Journal of the International Society of Cerebral Blood Flow and Metabolism
P A LiJ P MacManus

Abstract

Previous histopathologic results have suggested that one mechanism whereby hyperglycemia (HG) leads to exaggerated ischemic damage involves fragmentation of DNA. DNA fragmentation in normoglycemia (NG) and HG rats subjected to 30 minutes of forebrain ischemia was studied by terminal deoxynucleotidyl transferase mediated DNA nick-labeling (TUNEL) staining, by pulse-field gel electrophoresis (PFGE), and by ligation-mediated polymerase chain reaction (LM-PCR). High molecular weight DNA fragments were detected by PFGE, whereas low molecular weight DNA fragments were detected using LM-PCR techniques. The LM-PCR procedure was performed on DNA from test samples with blunt (without Klenow polymerase) and 3'-recessed ends (with Klenow polymerase). In addition, cytochrome c release and caspase-3 activation were studied by immunocytochemistry. Results show that HG causes cytochrome c release, activates caspase-3, and exacerbates DNA fragments induced by ischemia. Thus, in HG rats, but not in control or NGs, TUNEL-stained cells were found in the cingulate cortex, neocortex, thalamus, and dorsolateral crest of the striatum, where neuronal death was observed by conventional histopathology, and where both cytosolic cytochrome c and active cas...Continue Reading

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Citations

Mar 8, 2005·Journal of Neuroimmunology·Chaonan DingPing-An Li
Jul 12, 2007·Journal of Cerebral Blood Flow and Metabolism : Official Journal of the International Society of Cerebral Blood Flow and Metabolism·Xianren WuPatrick M Kochanek
Jan 29, 2008·Neuropharmacology·Konstantin-Alexander Hossmann
Nov 9, 2004·Life Sciences·Zhen-guo LiJoseph C Dunbar
Apr 30, 2013·Neurological Research·William A LiYuchuan Ding
Nov 3, 2011·Diabetes & Vascular Disease Research : Official Journal of the International Society of Diabetes and Vascular Disease·Lokesh K Bhatt, Veeranjaneyulu Addepalli
Jan 30, 2004·Diabetes Care·Stephen ClementUNKNOWN American Diabetes Association Diabetes in Hospitals Writing Committee

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Methods Mentioned

BETA
light microscopy
electrophoresis
PCR

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