Specific induction of protein kinase C delta subspecies after transient middle cerebral artery occlusion in the rat brain: inhibition by MK-801.
Protein kinase C (PKC) consists of a family of closely related Ca2+/phospholipid-dependent phosphotransferase isozymes, most of which are present in the brain and are differentially activated by second messengers. Calcium-dependent PKC activity may cause neuronal degeneration after ischemic insult. PKC is also involved in trophic-factor signaling, indicating that activity of some PKC subspecies may be beneficial to the injured brain. Therefore, we screened long-term changes in the expression of multiple PKC subspecies after focal brain ischemia. Middle cerebral artery occlusion was produced by using an intraluminal suture for 30 min of 90 min. In in situ hybridization experiments, mRNA levels of PKC alpha, -beta, -gamma, -delta, -epsilon, and -zeta were decreased in the infarct core 4 hr after ischemia and were lost completely 12 hr after ischemia. In areas surrounding the core, PKC delta mRNA was specifically induced 4, 12, and 24 hr after ischemia in the cortex. At 3 and 7 d, the core and a rim around it showed increased mRNA levels of PKC delta. No other subspecies were induced. At 2 d, immunoblotting demonstrated increased levels of PKC delta protein in the perifocal tissue, and immunocytochemistry revealed an increased number of PKC delta-positive neurons in the perifocal cortex. In the core, PKC delta-positive macrophages and endothelial cells were seen. Pretreatment with MK-801, an NMDA antagonist, inhibited cortical PKC delta mRNA induction. The data show that focal brain ischemia induces PKC delta mRNA and protein but not other PKC subspecies through the activation of NMDA receptors and that the upregulation lasts for several days in neurons of the perifocal zone.
Miettinen
S
S
A. I. Virtanen Institute, University of Kuopio, Finland.
Roivainen
R
R
Keinänen
R
R
Hökfelt
T
T
Koistinaho
J
J
D000818:Animals; D001921:Brain / Q000187:drug effects* / Q000201:enzymology* / Q000473:pathology; D016291:Dizocilpine Maleate / Q000494:pharmacology*; D004790:Enzyme Induction / Q000187:drug effects; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D002546:Ischemic Attack, Transient / Q000201:enzymology* / Q000473:pathology; D007527:Isoenzymes / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D008297:Male; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D051745:Protein Kinase C-delta; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D007527:Isoenzymes; D012333:RNA, Messenger; D016291:Dizocilpine Maleate; C497260:Prkcd protein, rat; D011493:Protein Kinase C; D051745:Protein Kinase C-delta