Elucidation of the mechanism of membrane transport, metallochaperone of essential trace metals related to central nervous activities

在本研究中，我们试图阐明影响中枢神经系统活动的痕量金属的代谢。在不同的培养细胞中，我们检测了铜的金属配位体CCS、从CCS接受铜的铜、锌-超氧化物歧化酶和铜的转运体CTR1在缺铜和过多状态下的变化。在缺铜状态下，CCS和铜、锌-超氧化物歧化酶的蛋白质含量增加。另一方面，在铜含量过高的状态下，CCS和铜、锌-超氧化物歧化酶的蛋白质含量显著下降。此外，我还让一只动物摄入了太多的锌。在锌含量过高的状态下，会出现肠道铜吸收梗阻。提示肠道细胞内铜含量减少，铜转运蛋白ATP7A也减少。在神经细胞来源的培养细胞中加入铜螯合物后，细胞内花生四烯酸代谢发生明显变化。铜缺乏导致铜、锌-超氧化物歧化酶活性下降，从而导致一氧化氮和活性氧的升高，从而影响细胞内花生四烯酸的代谢。我们检测了金属硫蛋白-3(MT-3)的金属结合能力。与其他金属硫蛋白相比，MT-3与铜的结合亲和力要高得多。通过神经传递的直观观察方法，我们阐明了三丁基锡对神经传递的影响。结果表明，三丁基锡阻碍了与学习记忆相关的长时程增强，并将引起中枢神经紊乱的金属结合蛋白与活性氧物种的障碍爆发机制进行了研究。

In the present study, we intended to clarify a metabolism of trace metals, which affected central nervous system activity. With various cultured cells, we examined changes of a metallochaperone of copper, CCS, Cu,Zn-SOD, which received copper from CCS, and Ctr1, which was copper transporter in a copper lack and too much states. In a copper lack state, the quantity of protein of CCS and Cu,Zn-SOD increased. On the other hand, in a state having too much copper, the quantity of protein of CCS and Cu,Zn-SOD showed a significant fall. Furthermore, I made an animal having too much a zinc intake. In a state having too much zinc, a copper absorption obstruction in an intestinal tract was occurred. It was suggested that the copper in intestinal tract cell decreased, and a copper transporter protein ATP7A decreased as well.When it added copper chelate in a cultured cell of a nerve cell origin, it became clear that arachidonic acid metabolism changed in a cell. A fall of Cu,Zn-SOD activity by copper lack caused rises of NO and active oxygen species as the cause and affected arachidonic acid metabolism in a cell.We examined metal binding ability of metallothionein-3 (MT-3), which specifically emerged to a central nervous system. It became clear that copper-binding affinity of MT-3 is very much higher in comparison with other metallothioneins.With a visual observation method of neurotransmission, we clarified influence of tributyltin on the neurotransmission. As a result, tributyltin obstructed the long-term potentiation, which was related to learning and memory.Furthermore, I put the metal-binding protein, which caused the central nervous disorder and examination of obstacle outbreak mechanism of active oxygen species.