Kouroku Y, Urase K, Fujita E, Isahara K, Ohsawa Y, Uchiyama Y, Momoi M Y, Momoi T
Division of Development and Differentiation, National Institute of Neuroscience, NCNP, Tokyo, Japan.
Biochem Biophys Res Commun. 1998 Jun 29;247(3):780-4. doi: 10.1006/bbrc.1998.8815.
We prepared a cleavage site-directed antiserum against Caspase-3 (anti-p20/17), which reacts with the p20/17 fragment (p20/17) activated by cleavage but not proCaspase-3 (p32), and examined the relationship between the activation of Caspase-3 and apoptosis. We identified p20/17-positive cells where cell death occurs naturally: interdigits of the forelimbs, small intestine epithelium, thymus, trigeminal ganglia, and dorsal root ganglia of mouse embryos. Withdrawal of nerve growth factor induced the appearance of p20/17-positive cells with DNA fragmentation in the culture of dorsal root ganglia neurons, while DNA fragmentation was detected in both p20/17-positive and -negative neurons in dorsal root ganglia of mouse embryos. These results suggest that not only activation of Caspase-3 but also other molecular mechanism play a role in the naturally occurring dorsal root ganglia apoptosis. Cleavage site-directed antisera against Caspases will be useful for the analysis of the molecular mechanism of naturally occurring apoptosis during development.
我们制备了一种针对半胱天冬酶-3的切割位点定向抗血清（抗-p20/17），它与经切割激活的p20/17片段（p20/17）反应，但不与前半胱天冬酶-3（p32）反应，并研究了半胱天冬酶-3的激活与细胞凋亡之间的关系。我们在自然发生细胞死亡的部位鉴定出p20/17阳性细胞：小鼠胚胎前肢的指间、小肠上皮、胸腺、三叉神经节和背根神经节。去除神经生长因子会在背根神经节神经元培养物中诱导出现具有DNA片段化的p20/17阳性细胞，而在小鼠胚胎背根神经节的p20/17阳性和阴性神经元中均检测到DNA片段化。这些结果表明，不仅半胱天冬酶-3的激活，而且其他分子机制在自然发生的背根神经节细胞凋亡中也起作用。针对半胱天冬酶的切割位点定向抗血清将有助于分析发育过程中自然发生的细胞凋亡的分子机制。
