Detection of the activation of caspases during neuronal cell death by using antiserum specifically recognizing active form of caspases

使用特异性识别半胱天冬酶活性形式的抗血清检测神经元细胞死亡过程中半胱天冬酶的激活

• 批准号: 11480235
• 负责人: MOMOI Takashi
• 金额: $ 7.55万
• 依托单位: National Institute of Neuroscience
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11480235/
• 关键词: apoptosis; caspase-12; ER stress; polyglutamine disease; Huntinton; Endoplasmic Reticulum; processing; Neurodegenerative disease; カスパーゼー12; ER ストレス; ハンチントン病; ポリグルタミン; カスパーゼ12; 神経細胞死; 脳形成; カスパーゼ9; カスパーゼ3; Bcl-x; ミトコンドリア; チトクロームC; Akt

Considerable evidence suggests that caspase, a member of cystein protease, plays an essential role in the various cell death including apoptosis and programmed cell death (PCD) and in the cell death observed in the various diseases. However, there is little direct evidence on the involvement of caspases in naturally occurring cell death during development of the vertebrate nervous system and diseases. Caspases are sequentially activated in the caspase cascade, in which activated initial caspases such as caspase 9, caspase 8 and caspase 12 are autoprocessed and then process caspase 3, 6 and 7. To clarify the involvement of caspases in the PCD and neurodegenerative diseases, we prepared cleavage site-directed antiserum against 12 members of caspases that specifically react with the cleaved fragment of caspases, respectively. We have examined directly the activation of caspases during development of mouse embryos and subcellular localization of active form of caspases by immunohistochemical staining using these antisera.

大量证据表明半胱氨酸蛋白酶的成员之一的半胱天冬酶在包括细胞凋亡和程序性细胞死亡（PCD）在内的各种细胞死亡以及在各种疾病中观察到的细胞死亡中发挥重要作用。然而，几乎没有直接证据表明半胱天冬酶参与脊椎动物神经系统和疾病发育过程中自然发生的细胞死亡。 Caspase 在 Caspase 级联中依次被激活，其中激活的初始 Caspase，如 Caspase 9、Caspase 8 和 Caspase 12 被自动处理，然后处理 Caspase 3、6 和 7。为了阐明 Caspase 在 PCD 和神经退行性疾病中的参与，我们制备了针对 12 个半胱天冬酶成员的切割位点定向抗血清，这些成员与 caspase 的切割片段特异性反应分别是半胱天冬酶。我们使用这些抗血清通过免疫组织化学染色直接检查了小鼠胚胎发育过程中半胱天冬酶的激活以及活性半胱天冬酶的亚细胞定位。

项目成果

Kouroku, Y., Fujita, E., Jimbo, A., Kikuchi, T., Yamagata, T., Momoi, M..Y., Kominami, E., Kuida, K., Sakamaki, K., Yonehara, S., and Momoi, T.: "Polyglutamine aggregates stimulate ER stress signals and caspase-12 activation"Human Mol. Genet. 11. 1505-151

Kouroku，Y.，藤田，E.，Jimbo，A.，菊池，T.，山形，T.，桃井，M..Y.，小南，E.，Kuida，K.，坂卷，K.，米原，

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Jimbo, A.、Fujila, B.、Kouroku, Y.、Ohnishi, J.、Inohara, N.、Kuida, K.、Sakainaki, K.、Yonehara.S.、Momoi, T.：“ER 应激诱导 caspase