一氧化氮是高钾氯化钾去极化诱导PC12细胞分化的基础。

Nitric oxide underlies the differentiation of PC12 cells induced by depolarization with high KCl.

作者信息

Nakagawa H, Yoshida M, Miyamoto S

机构信息

Department of Biochemical Science, Faculty of Computer and Systems, Engineering, Kyushu Institute of Technology, Iizuka, Fukuoka 820-8502, Japan.

出版信息

J Biochem. 2000 Jan;127(1):113-9. doi: 10.1093/oxfordjournals.jbchem.a022571.

DOI:10.1093/oxfordjournals.jbchem.a022571

PMID:10731673

Abstract

Nitric oxide (NO) acts as a cytostatic agent to induce neuronal differentiation of PC12 cells after nerve growth factor (NGF) treatment. We newly subcloned PC12K cells that extended neurites after depolarization with high KCl. Here we present evidence that the neuronal differentiation of PC12K cells caused by depolarization with high KCl is mediated by endogenous NO. The outgrowth of neurites was significantly inhibited by 2 mM N-nitro-L-arginine methyl ester (L-NMAE), and 10 mM L-NAME was necessary for complete inhibition. The inhibition of NGF-dependent neurite outgrowth by L-NAME was abolished by depolarization of cells with KCl. The expression of neuronal- and endothelial-NO-synthase in PC12K cells was confirmed by immuno-cytochemical and immuno-blotting analyses with the respective monoclonal antibodies. However, the expression of inducible-NO synthase was not observed in PC12K cells cultured with high KCl under the depolarization conditions with 45 mM KCl. We observed the increase of NO in the differentiated PC12K cells using diaminofluorescein, a novel fluorescent indicator for NO.

摘要

一氧化氮（NO）在神经生长因子（NGF）处理后作为一种细胞生长抑制剂诱导PC12细胞的神经元分化。我们新亚克隆了PC12K细胞，该细胞在用高浓度氯化钾去极化后会延伸神经突。在此我们提供证据表明，高浓度氯化钾去极化所导致的PC12K细胞的神经元分化是由内源性NO介导的。神经突的生长被2 mM N-硝基-L-精氨酸甲酯（L-NMAE）显著抑制，而完全抑制则需要10 mM L-NAME。用氯化钾使细胞去极化可消除L-NAME对NGF依赖性神经突生长的抑制作用。通过使用各自的单克隆抗体进行免疫细胞化学和免疫印迹分析，证实了PC12K细胞中神经元型和内皮型一氧化氮合酶的表达。然而，在45 mM氯化钾去极化条件下用高浓度氯化钾培养的PC12K细胞中未观察到诱导型一氧化氮合酶的表达。我们使用二氨基荧光素（一种新型的NO荧光指示剂）观察到分化的PC12K细胞中NO的增加。