Post-ischemic changes in protein kinase C RNA in the gerbil brain following prolonged periods of recirculation: a phosphorimaging study.

Abstract

Northern blot analysis was performed to investigate the long-term changes in mRNA expression of protein kinase C (PKC) in the gerbil brain following transient cerebral ischemia. We have previously demonstrated an increase in mRNA levels of certain Ca(2+)-independent forms of PKC in early recirculation periods i.e., 6 h and 24 h postischemia (PI). But, since neuronal death in susceptible regions usually occurs 2-3 days following ischemia, this study examined the mRNA levels of PKC after prolonged periods of reperfusion following ischemia. The mRNA expression was also examined at an early recirculation period, i.e., 1 h, to determine how early the alterations begin to occur. Global forebrain ischemia was produced in gerbils by 10 min of bilateral carotid artery occlusion. RNA was prepared from forebrains of nonischemic controls and PI animals following 1 h, 3 d, and 7 d of recirculation (n = 3 to 4 in each group) and hybridized with synthetic oligonucleotide probes for PKC, delta, epsilon, and zeta based on cDNA sequences in rat and labelled with 32P. The autoradiographs were recorded and quantified by a sensitive system, Phosphor Imager, followed by conventional x-ray film exposure. The mRNA levels of all 3 PKC isozymes examined were found to be elevated as early as 1 h recirculation following ischemia. The increases in mRNA levels of both delta PKC following 6 h and 24 h of recirculation as well as that of zeta PKC following 24 h of recirculation, as reported earlier, return to control levels by 3 d PI and remain at that level 7 d PI.(ABSTRACT TRUNCATED AT 250 WORDS)