蛋白激酶C在PC12细胞中二氧化碳/氢离子诱导的c-fos信使核糖核酸表达中的可能作用。

A possible role for protein kinase C in CO2/H+-induced c-fos mRNA expression in PC12 cells.

作者信息

Kuo N T, Agani F H, Haxhiu M A, Chang C H

机构信息

Case Western Reserve University, Cleveland, Ohio 44106, USA.

出版信息

Respir Physiol. 1998 Feb;111(2):127-35. doi: 10.1016/s0034-5687(97)00115-1.

DOI:10.1016/s0034-5687(97)00115-1

PMID:9574865

Abstract

Recently we have found that hypercapnia induces nuclear protein (FOS) expression in the brainstem chemosensitive neurons, including catecholamine-containing cells. In the present studies we examined the role of protein kinase C (PKC) pathway in CO2-induced c-fos expression. Because of the complexity of the CNS system, experiments were performed in pheochromocytoma cells (PC12 cells). These cells originate from neuronal crest and express catecholaminergic traits. We depleted PKC from PC12 cells by prolonged (48 h) exposure to high concentration of phorbol 12-myristate, 13-acetate (PMA, 100 nM), and then determined the expression of: (1) c-fos mRNA by Northern blot (2) PKC isoforms, tyrosine phosphorylated and unphosphorylated MAP (mitogen activated protein) kinases by Western blot. Depletion of PKC abolished the effect of CO2 on c-fos mRNA expression, inhibited MAP kinases tyrosine phosphorylation and suppressed the expression of PKC(alpha) and PKC(zeta). These results suggest that MAP kinases, PKC(alpha) and/or PKC(beta) might be involved in CO2-induced c-fos mRNA expression.

摘要

最近我们发现，高碳酸血症可诱导脑干化学敏感神经元（包括含儿茶酚胺的细胞）中的核蛋白（FOS）表达。在本研究中，我们检测了蛋白激酶C（PKC）通路在二氧化碳诱导的c-fos表达中的作用。由于中枢神经系统的复杂性，实验在嗜铬细胞瘤细胞（PC12细胞）中进行。这些细胞起源于神经嵴并表达儿茶酚胺能特性。我们通过长时间（48小时）暴露于高浓度佛波酯12-肉豆蔻酸酯、13-乙酸酯（PMA，100 nM）来消耗PC12细胞中的PKC，然后通过Northern印迹法测定（1）c-fos mRNA的表达，通过蛋白质印迹法测定（2）PKC亚型、酪氨酸磷酸化和未磷酸化的丝裂原活化蛋白（MAP）激酶。PKC的消耗消除了二氧化碳对c-fos mRNA表达的影响，抑制了MAP激酶的酪氨酸磷酸化，并抑制了PKC（α）和PKC（ζ）的表达。这些结果表明，MAP激酶、PKC（α）和/或PKC（β）可能参与了二氧化碳诱导的c-fos mRNA表达。