title: Transglutaminase activity and transglutaminase mRNA transcripts in gerbil brain ischemia.
abstract: Brain injury of the ischemia/reperfusion type induces neuronal damage, mainly by excitatory amino acid release, intracellular Ca(2+) overload and reactive oxygen species production. We have previously demonstrated that glutamate exposure increased transglutaminase activity and transglutaminase 2 expression in cultured cerebellar granule cells and astrocytes. The aim of this study is to evaluate changes in transglutaminase activity and expression using a gerbil model of global cerebral ischemia. Moreover, the distribution and amounts of different transglutaminase isoforms were examined. Transglutaminase activity was measured by incorporation of [(3)H]putrescine into dimethylcasein throughout 48 h of reperfusion following a 3 min occlusion. Compared to sham-operated brains, significant increases were found in the ischemic hippocampus at 24 h of reperfusion, while minor changes were observed in the cortex. RT-PCR demonstrated the presence of significant mRNA amounts of transglutaminase 2 and transglutaminase 1, both in the hippocampus and the cerebral cortex, while low levels were found for transglutaminase 3 transcripts. Interestingly, transglutaminase 2 and transglutaminase 1 mRNAs were 4-fold and 2-fold increased, respectively, in the ischemic hippocampus after 24 h of reperfusion. Western blot analysis of transglutaminase 2 expression confirmed a strong up-regulation in the ischemic hippocampus. However, it is possible to hypothesize that different expression rates of transglutaminase isoforms may be dependent on different responsiveness of their transcription regulatory elements to intracellular calcium overload following excitotoxic cell injury. Our results suggest that increases in transglutaminases may be part of the tissue stress response in global brain ischemia.
authors.affiliation: Department of Biochemical, Physiological and Nutritional Sciences, University of Messina, Messina, Italy. ientile@unime.it
authors.forename: Riccardo
authors.initials: R
authors.lastname: Ientile
authors.forename: Daniela
authors.initials: D
authors.lastname: Caccamo
authors.forename: Maria Concetta
authors.initials: MC
authors.lastname: Marciano
authors.forename: Monica
authors.initials: M
authors.lastname: Currò
authors.forename: Carmen
authors.initials: C
authors.lastname: Mannucci
authors.forename: Agata
authors.initials: A
authors.lastname: Campisi
authors.forename: Gioacchino
authors.initials: G
authors.lastname: Calapai
mesh_terms: D000818:Animals; D001921:Brain / Q000098:blood supply / Q000201:enzymology* / Q000503:physiopathology; D002545:Brain Ischemia / Q000201:enzymology* / Q000235:genetics* / Q000503:physiopathology; D020013:Calcium Signaling / Q000235:genetics; D002540:Cerebral Cortex / Q000201:enzymology / Q000503:physiopathology; D004195:Disease Models, Animal; D005809:Genes, Regulator / Q000235:genetics; D005849:Gerbillinae; D018698:Glutamic Acid / Q000378:metabolism; D006624:Hippocampus / Q000201:enzymology / Q000503:physiopathology; D002546:Ischemic Attack, Transient / Q000201:enzymology / Q000235:genetics / Q000503:physiopathology; D008297:Male; D009498:Neurotoxins / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D015427:Reperfusion Injury / Q000201:enzymology / Q000235:genetics / Q000503:physiopathology; D011503:Transglutaminases / Q000235:genetics* / Q000378:metabolism*; D015854:Up-Regulation / Q000235:genetics
chemical_list: D009498:Neurotoxins; D020033:Protein Isoforms; D012333:RNA, Messenger; D018698:Glutamic Acid; D011503:Transglutaminases