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（caspase）是半胱氨酸蛋白酶的成员，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起重要作用。然而，几乎没有直接证据表明，在脊椎动物神经系统和疾病的发展过程中，半胱天冬酶参与自然发生的细胞死亡。半胱天冬酶在半胱天冬酶级联中顺序活化，其中活化的初始半胱天冬酶如半胱天冬酶9、半胱天冬酶8和半胱天冬酶12被自动加工，然后加工半胱天冬酶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caspase成员的切割位点定向抗血清，其分别与caspase的切割片段特异性反应。我们直接研究了小鼠胚胎发育过程中半胱天冬酶的激活和半胱天冬酶活性形式的亚细胞定位，免疫组化染色使用这些抗血清。

项目成果

Kitanaka C et al.: "Increased ras expression and caspase-independent neuroblastoma cell death : possible mechanism of spontaneous neuroblastoma regression."J Natl Cancer Inst.. 94. 358-368 (2002)

Kitanaka C 等人：“ras 表达增加和不依赖 caspase 的神经母细胞瘤细胞死亡：自发性神经母细胞瘤消退的可能机制。”J Natl Cancer Inst.. 94. 358-368 (2002)

Shigemura N et al.: "Localization of activated caspase-3-positive and apoptotic cells in the developing tooth germ of the mouse lower first molar"Histochem J.. 33. 253-258 (2001)

Shigemura N 等人：“小鼠下第一磨牙发育中的牙胚中活化的 caspase-3 阳性细胞和凋亡细胞的定位”Histochem J.. 33. 253-258 (2001)

Takahashi, K., Kamiya, K., Urase, K., Fujita, E., Suga, M., Mori, H., Yoshikawa, Y., Ichimura, K., Kuida, K., Momoi, T.: "Caspase-3-deficiency induces hyperplasia of supporting cells and degeneration of sensory cells resulting in the hearing loss"Brain Re

高桥 K.、神谷 K.、浦濑 K.、藤田 E.、须贺 M.、森 H.、吉川 Y.、市村 K.、久田 K.、桃井 T.：

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，

Fujita, E. et al.: "BMP-4 and retinoic acid synergistically induce activation of caspase-9 and cause apoptosis of P19 embryonal carcinoma cells cultured as a monolayer"Cell Death Differ.. 6. 1109-1116 (1999)

Fujita, E. 等人：“BMP-4 和视黄酸协同诱导 caspase-9 的激活并导致单层培养的 P19 胚胎癌细胞凋亡”细胞死亡差异.. 6. 1109-1116 (1999)