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

大量证据表明，半胱氨酸蛋白酶的成员caspase在包括凋亡和程序性细胞死亡（PCD）在内的各种细胞死亡以及各种疾病中观察到的细胞死亡中起着重要作用。然而，很少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级联中被顺序激活，其中激活的初始caspase如caspase 9、caspase 8和caspase 12被自动加工，然后加工caspase 3、6和7。为了阐明caspases在PCD和神经退行性疾病中的作用，我们制备了针对12个caspases成员的靶向抗血清，这些caspases成员分别与caspases被切割的片段发生特异性反应。我们使用这些抗血清进行免疫组化染色，直接检测了小鼠胚胎发育过程中半胱氨酸酶的激活和半胱氨酸酶活性形式的亚细胞定位。

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.