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Effects of extracellular ATP and nerve growth factor on pheochromocytoma PC12 cells.

细胞外ATP和神经生长因子对嗜铬细胞瘤PC12细胞的影响。

基本信息

批准号：
01571027
负责人：
YAMADA Shozo
金额：
$ 1.41万
依托单位：
Meikai University School of Dentistry
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01571027/
关键词：
Extracellular ATP Pheochromocytoma PC12 Nerve growth factor Intracellular calcium ion Cyclic AMP P_2 purinergic receptor Catecholamine release Dopamine production カルシウムチャンネル神経細胞分化神経突起形成

项目摘要

Effects of extracellular ATP on pheochrowxytoma PC12 and PC12h cells, and on the responses of the cells to nerve growth factor (NGF) were studied. 1) ATP inhibited the PC12h cell growth regardless with the presence or absence of NGF in the culture medium. 2) ATP enhanced the NGF-induced neurite outgrowth from PC12h cells : it increased the neurite length rather than the frequency of the neurite formation. ADP, AMP and adenosine (ADO) also showed the similar effect. Rank order of their potency was ATP>ADP>AMP=ADO. 3) ATP stimulated phosphorylation of a nuclear protein, SMP, of PC12 cells. SMP is a protein of which phosphorylation has been known to be enhanced by NGF. At lower concentration this enhancing effect was specific for ATP. At higher concentration (1 mM) AMP and ADO also showed the similar effect. 4) ATPinduced increase in cytosolic Ca^<2+> concentration of Pcl2h cells. Even in the absence of extracellular Ca^<2+>, about 40% of the maximal[Ca^<2+>]i increase was induced by ATP. ADP had the similar effect but AMP and ADO did not. 5) ATP induced a release of dopamine from PC12h cells. In the absence of extracellular Ca^<2+>, less than 10% of dopamine release detected in the presence of Ca^<2+> was observed, indicating that Ca^<2+> introduced into the cells by influx may play the major role in ATP-evoked catecholamine release. In the short term responses of the cells to ATP, adenine nucleotide specificity for ATP was quite clear. On the other hand, ATP is known to metabolized to ADP, AMP and ADO in sermcontaining culture medium and in body fluid. Therefore, the responses of the cells observed by long-term treatment with ATP may be resulted from a complex effect of ATP mediated by P_2 receptors and that of AMP and ADO mediated by Adenosine receptors.

研究了细胞外ATP对PC12和PC12h细胞的作用，以及细胞对神经生长因子(NGF)的反应。1)无论培养液中有无NGF，ATP均能抑制PC12h细胞的生长。2)ATP促进NGF诱导的PC12h细胞突起生长：增加突起的长度，而不是增加突起的形成频率。腺苷二磷酸(ADP)、腺苷(AMP)和腺苷(ADO)也表现出类似的作用。其效价顺序为：ATP&GT；ADP&GT；AMP=ADO。3)ATP刺激PC12细胞核蛋白SMP的磷酸化。SMP是一种已知的被NGF增强其磷酸化的蛋白质。在较低浓度时，这种增强作用是ATP所特有的。在较高浓度(1 MM)时，AMP和ADO也表现出类似的作用。(4)ATP诱导Pcl2 h细胞内游离钙离子浓度升高。即使在无细胞外[Ca~(2+)]i的情况下，约有40%的最大[Ca~(2+)]i升高是由ATP引起的。ADP有类似的作用，而AMP和ADO则不起作用。5)ATP可诱导PC12h细胞释放多巴胺。在没有胞外Ca~(2+)的情况下，观察到只有不到10%的多巴胺在有Ca~(2+)的存在下被释放，这表明通过内流进入细胞内的Ca~(2+)可能在ATP引起的儿茶酚胺释放中起主要作用。在细胞对三磷酸腺苷的短期反应中，腺嘌呤核苷酸对三磷酸腺苷的特异性非常明显。另一方面，在含血清的培养液和体液中，ATP被代谢成ADP、AMP和ADO。因此，长期用三磷酸腺苷处理细胞的反应可能是由P_2受体介导的三磷酸腺苷和腺苷受体介导的腺苷和腺苷共同作用的结果。