Imaging and functional regulation of microenvironment in subcellular organelle

亚细胞器微环境的成像和功能调控

基本信息

批准号：
15390064
负责人：
MATSUI Hideki
金额：
$ 9.86万
依托单位：
OKAYAMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15390064/
关键词：
Calcineurin Calpain Calpastatin Neuronal cell death Glutamate Kainate FK506 Protein therapy

项目摘要

Excessive glutamate as an excitotoxic inducer plays an important role in acute and chronic neuronal cell death including ischemia, trauma, Huntington's disease, Alzheimer's disease, and Amyotrophic Lateral Sclerosis. Despite years of investigation, the precise mechanism involved in glutamate-induced excitotoxic neuronal cell death has not been well characterized.Calcineurin(CaN) and calpain, a Ca^<2+>/calmodulin (CaM)-dependent protein phosphatase and a Ca^<2+>-dependent cysteine protease, respevtively, are thought to be two of the most important Ca^<2+>-dependent effectors involved in neuronal cell death. Although the mechanism of CaN-mediated neuronal cell death remains controversial, one proposed mechanism that calcineurin triggers dysfunction of the mitochondria in degenerative neurons has been strongly supportedOn the other hand, calpain regulates physiological signal transduction by cleaving cytoskeletal proteins, membrane proteins and enzymes and is also thought to play a critic … More al role in a form of neuronal cell death involved in the pathogenesis of several diseases such as brain injury, Alzheimer's disease and focal or global cerebral ischemia. Some substrates of calpain such as p35, a specific activator of cyclin-dependent kinase 5(Cdk5) have been suggested as key molecules in the induction of neuronal cell death.In this research, we showed that during glutamate neuroexcitotoxicity, calcineurin A(CnA) is directly cleaved by calpain in vitro and in vivo, resulting in the enzyme being converted to an active form. Mass spectrometry identified the three cleavage sites in CnA and two of them were constitutively active forms. Overexpression of the cleaved CnA induced caspase activity and neuronal cell death. Calpain inhibitors not only blocked the cleavage of calcineurin, but also protected against neuronal cell death in vitro and in vivo. These results indicate that calcineurin is a crucial target for calpain and the calpain-mediated activation of calcineurin triggers excitotoxic neurodegeneration. This further shows the direct cross talk between phosphatase and protease in pathopysiology. Less

过度的谷氨酸作为兴奋性毒性诱导在急性和慢性神经元细胞死亡中起重要作用，包括缺血，创伤，亨廷顿氏病，阿尔茨海默氏病和肌萎缩性侧面硬化症。尽管进行了多年的研究，但谷氨酸诱导的兴奋性神经元细胞死亡中涉及的精确机制尚未得到很好的特征。CALCINERIN（CAN）和CALPAIN，Ca^<2+>/钙调蛋白（CAM）依赖性蛋白磷酸酶以及Ca^<2+> - 依赖性的pectectectionding a pectiNTICE <2神经元细胞死亡涉及的作用。 Although the mechanism of CaN-mediated neuronal cell death remains controversial, one proposed mechanism that calcineurin triggers dysfunction of the mitochondria in degenerative neurons has been strongly supported On the other hand, calpain regulates physical signal transduction by Cleaving cytoskeletal proteins, membrane proteins and enzymes and is also thought to play a critical … More al role in a form of神经元细胞死亡涉及多种疾病的发病机理，例如脑损伤，阿尔茨海默氏病以及局灶性或全球脑缺血。在诱导神经元细胞死亡的诱导中，已经提出了一些钙蛋白酶（例如p35）的层底物，例如p35，p35，p35的特异性激活剂，这是在这项研究中作为关键分子。活跃形式。质谱法确定了CNA中的三个切割位点，其中两个始终是活跃的形式。切割CNA的过表达诱导caspase活性和神经元细胞死亡。钙蛋白酶抑制剂不仅阻塞了钙调蛋白的裂解，而且还可以保护体外和体内神经元细胞死亡。这些结果表明，钙调神经蛋白是钙蛋白酶的关键靶标，而钙调蛋白介导的钙调神经磷酸酶的激活触发了兴奋性神经变性。这进一步显示了病原体生理学中磷酸酶和蛋白酶之间的直接串扰。较少的