Expression of protein kinase C in postischemic brain: an in situ hybridization study.
Cerebral ischemia leads to a number of biochemical and molecular changes which include increase in intracellular calcium, arachidonic acid, and diacylglycerol, all of which are capable of activating protein kinase C (PKC). To investigate how the expression of PKC is affected in postischemic brain, ischemia was produced in gerbils by bilateral common carotid artery occlusion for 10 min followed by reperfusion for 15 min, 6 h, and 24 h. The brains of postischemic and normal control animals were removed, forebrains dissected, fresh frozen, and processed for in situ hybridization. The mRNA expression of PKC was analyzed by using oligonucleotide probes based on the sequences of PKC alpha and epsilon isozymes in this preliminary study. There was no change observed in the expression of PKC alpha in any region of the brain in any of the postischemic groups examined. There was, however, a qualitative increase in the transcription for PKC epsilon in two out of three brains of 15 min postischemic group which continued through 24 h of reperfusion. Since the protein itself was not examined, it can not be said how these observations regarding transcription relate to the synthesis of the protein and whether there are any changes in the subcellular distribution of PKC following ischemia. However, since there was no decrease in transcription demonstrated in our study, it appears that the reported decrease in PKC activity following ischemia is not due to decreased mRNA expression.
Kumar
K
K
Department of Pathology, Michigan State University, East Lansing 48824.
Savithiry
S
S
Madhukar
B
B
D000818:Animals; D001921:Brain / Q000098:blood supply* / Q000201:enzymology; D005849:Gerbillinae; D007511:Ischemia / Q000201:enzymology*; D007527:Isoenzymes / Q000032:analysis; D008297:Male; D009693:Nucleic Acid Hybridization; D011493:Protein Kinase C / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis*; D014158:Transcription, Genetic
D007527:Isoenzymes; D012333:RNA, Messenger; D011493:Protein Kinase C