Freire-Moar J, Cherwinski H, Hwang F, Ransom J, Webb D
Department of Molecular Immunology, Syntex Research, Palo Alto, CA 94304.
J Immunol. 1991 Jul 15;147(2):405-9.
The expression of the different protein kinase C (PKC) isozymes in mouse thymocytes was studied to determine if there is a correlation between isozyme expression and thymocyte phenotype. Expression of PKC isozymes in thymocyte subsets (distinguished by the CD4 or CD8 Ag) was determined by message amplification phenotyping. The expression of mRNA for PKC-alpha, -beta, -epsilon, and -zeta, but not -gamma or -delta isozymes, was detected in all of the unstimulated thymocyte subpopulations analyzed. Thus no differences in the pattern of PKC isozyme expression were found that could be correlated with thymocyte phenotype. However, it was noted that the levels of PKC mRNA expression were affected by different stimuli in unfractionated thymocytes. Whereas mRNA levels of PKC-alpha and -beta were down-regulated by PMA and ionomycin treatment, no significant changes were seen in the levels of PKC-epsilon mRNA with these agents. PKC-epsilon mRNA decreased in thymocytes exposed to Con A similar to what has been reported for PKC-epsilon protein. PKC-zeta mRNA was also down-regulated by PMA or ionomycin, and the combination of both compounds caused a more rapid and drastic effect. Finally, PKC-delta mRNA expression was induced transiently in thymocytes only after exposure to PMA or Con A, and this induction was inhibited by ionomycin treatment. These results indicate that message levels of specific isoforms of PKC are uniquely regulated and suggest an additional level of control of PKC activity in activated lymphocytes.
研究了小鼠胸腺细胞中不同蛋白激酶C(PKC)同工酶的表达,以确定同工酶表达与胸腺细胞表型之间是否存在相关性。通过信息扩增表型分析确定胸腺细胞亚群(由CD4或CD8抗原区分)中PKC同工酶的表达。在所有分析的未刺激胸腺细胞亚群中均检测到PKC-α、-β、-ε和-ζ的mRNA表达,但未检测到PKC-γ或-δ同工酶的表达。因此,未发现PKC同工酶表达模式的差异与胸腺细胞表型相关。然而,值得注意的是,未分离的胸腺细胞中PKC mRNA表达水平受不同刺激的影响。PKC-α和-β的mRNA水平在佛波酯(PMA)和离子霉素处理后下调,而这些试剂处理后PKC-ε mRNA水平未见明显变化。与PKC-ε蛋白的报道情况类似,暴露于刀豆蛋白A(Con A)的胸腺细胞中PKC-ε mRNA减少。PKC-ζ mRNA也被PMA或离子霉素下调,两种化合物联合使用产生更快速、更显著的效果。最后,仅在暴露于PMA或Con A后,PKC-δ mRNA表达在胸腺细胞中短暂诱导,且这种诱导被离子霉素处理抑制。这些结果表明,PKC特定同工型的信息水平受到独特调控,并提示在活化淋巴细胞中PKC活性存在额外的调控水平。