Identification of P2-purinergic receptors

P2-嘌呤能受体的鉴定

基本信息

批准号：
09672208
负责人：
MURAYAMA Toshihiko
金额：
$ 1.28万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672208/
关键词：
ATP receptor PC12 cells glial cells adenylyl cyclase ARF S-nitroso-cysteine NO synthase ATP プリン受容体 GTP結合蛋白質 NO ホスホリパーゼA_2 アラキドン酸

项目摘要

1) PC12 pheochromocytoma cells have P2 receptors which are coupled to Ca2+influx and catecholamine release. In the presence of forskolin, an activator of adenylyl cyclase, ATP analogs such as ATP and 2-methylthio ATP inhibited cyclic AMP accumulation in a concentration-dependent manner. Treatment with pertussis toxin, which completely abolished the effect of carbachol, had no effect on the action of ATP.In addition, we found that ADP-ribosylation factors translocate to membranes from the cytosol fraction after exocytotic stimulation. The cloning of a new type of ATP receptor remains to be determined.2) Nitric oxide (NO) modulates the release of neurotransmitters. Previously we reported that 5-nitroso-cysteine stimulated noradrenaline release from hippocampus in vivo and in vitro. In PC12 cells, S-nitroso-cysteine did inhibit noradrenaline release. Other NO compounds, which increased cyclic GMP accumulation, had no effect. ATP-stimulated Ca2+ influx via Ca2+ channels was inhibited in PC12 cells treated with S-Nitroso-cysteine, although S-nitroso-cysteine stimulated Ca2+ mobilization from intracellular caffeine-sensitive Ca2+-pools. Ca2+ mobilization by NO from Ca2+ pools was not a sufficient factor, and other factors stimulating release may be regulated negatively.3) Lipopolysaccaride or cytokines are known to stimulate production of nitrite via expression of inducible NO synthase (iNOS) in rat glial cells. Co-addition of endothelin decreased iNOS expression and nitrite accumulation by stimulants. In contrast, pretreatment with endothelin or ATP for 24 h enhanced iNOS expression. The stimulatory effect by endothelin or ATP was mediated by ET-B or P2 receptors, respectively. A protein kinase C inhibitor suppressed endothelin- and ATP-enhanced iNOS expression. Stimulation of ATP receptors induced activation of a nuclear factor, NFkB in glial cells.

1）PC12嗜铬细胞瘤细胞的P2受体与Ca2+流入和儿茶酚胺释放耦合。在存在腺苷酸环化酶激活剂Forskolin的情况下，ATP类似物（例如ATP和2-甲基硫硫代）以浓度依赖性方式抑制环状AMP的积累。百日咳毒素的治疗完全消除了卡尔巴醇的作用，对ATP的作用没有影响。加上ADP-核糖基化因子在胞外刺激后从细胞质分数转移到膜上的ADP-核糖基化因子。新型ATP受体的克隆仍有待确定。2）一氧化氮（NO）调节神经递质的释放。以前，我们报道说，5-硝基半胱氨酸在体内和体外刺激了从海马释放去甲肾上腺素的释放。在PC12细胞中，S-亚硝基结晶性确实抑制了去甲肾上腺素的释放。没有增加环状GMP积累的其他化合物没有作用。在用S-硝基 - 半胱氨酸处理的PC12细胞中抑制了ATP刺激的Ca2+流入，尽管S-硝基 - 半胱氨酸刺激了细胞内咖啡因敏感的Ca2+ -Pools的Ca2+动员。 Ca2+从Ca2+池中的NO动员不够的因素，这不是足够的因素，其他刺激释放的因素可能受到负调节。3）已知脂多多藻酸或细胞因子通过在大鼠神经胶质细胞中表达诱导型NO合酶（Inos）来刺激亚硝酸盐的产生。内皮素的共同添加降低了iNOS表达和亚硝酸盐的积累。相反，用内皮素或ATP预处理24小时，增强了iNOS表达。内皮素或ATP的刺激作用分别由ET-B或P2受体介导。蛋白激酶C抑制剂抑制了内皮素 - 增强iNOS的表达。刺激ATP受体诱导的核因子激活，胶质细胞中的NFKB。