Up-regulation of tissue-type transglutaminase after traumatic brain injury.
Tissue-type transglutaminase (tTG, EC 2.3.2.13) has been implicated in various disease paradigms including neurodegenerative disease. In these studies, tTG induction after traumatic brain injury was studied using a rat cortical impact model. Using western blots, two forms of tTG protein expression were identified--an approximately 79-kDa primary form (tTG-L) and a less abundant approximately 70-kDa form (tTG-S). Both forms of tTG protein were elevated after injury. In ipsilateral cortex, peak induction of tTG-L protein [561% +/- 80% of control (n = 5)] was observed five days after injury, with expression remaining elevated after two weeks. Peak induction of tTG-S protein [302% +/- 81% of control (n = 5)] was observed three days after injury. Lesser tTG protein induction was observed in hippocampus. Northern blot analysis demonstrated two tTG transcripts in the ipsilateral cortex with peak induction of tTG-L mRNA three days after injury. However, tTG-S mRNA was not identified in control samples and only faintly detected in injured tissue. To facilitate analysis of low abundance transcripts in smaller tissue samples, a semiquantitative real-time PCR strategy was used. Semi-quantitative PCR analysis of tTG-L mRNA induction in ipsilateral cortex (peak after three days; 414% +/- 21% of control, n = 3) confirmed tTG-L mRNA induction determined by northern blot (410% of control).
Tolentino
Paul J
PJ
Department of Neuroscience, Evelyn F. and William L. McKnight Brain Institute, University of Florida Center for Traumatic Brain Injury Studies, Gainesville, Florida, USA.
DeFord
S Michelle
SM
Notterpek
Lucia
L
Glenn
Christopher C
CC
Pike
Brian R
BR
Wang
Kevin K W
KK
Hayes
Ronald L
RL
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D001930:Brain Injuries / Q000201:enzymology*; D002138:Calibration; D002540:Cerebral Cortex / Q000201:enzymology / Q000293:injuries; D004195:Disease Models, Animal; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D006624:Hippocampus / Q000201:enzymology / Q000293:injuries; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D016014:Linear Models; D008297:Male; D016133:Polymerase Chain Reaction; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011503:Transglutaminases / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation
D007527:Isoenzymes; D012333:RNA, Messenger; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins