Antibody to caspase-cleaved actin detects apoptosis in differentiated neuroblastoma and plaque-associated neurons and microglia in Alzheimer's disease.
Antibody to caspase-cleaved actin detects apoptosis in differentiated neuroblastoma and plaque-associated neurons and microglia in Alzheimer's disease.
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发表时间：
1998-02
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通讯作者：
Fusheng Yang;Xiaoyan Sun;W. Beech;B. Teter;Sherry Wu;J. Sigel;H. Vinters;S. Frautschy;G. Cole
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作者：
Fusheng Yang;Xiaoyan Sun;W. Beech;B. Teter;Sherry Wu;J. Sigel;H. Vinters;S. Frautschy;G. Cole
在细胞凋亡过程中，与白细胞介素-1 β转化酶（ICE）相关的蛋白酶或“半胱天冬酶”相关的半胱氨酸蛋白酶家族的激活导致多种底物在特定天冬氨酸残基处的内切蛋白水解裂解。我们已经寻求开发新的抗体探针的ICE相关的蛋白水解裂解产生的蛋白质片段中的新表位作为与凋亡机制紧密相连的事件的特异性标志物。在这里，我们证明了特异性的抗体探针的C末端的32 kd肌动蛋白片段产生的ICE样活性特异性标签凋亡，但不坏死，分化的人神经母细胞瘤细胞培养。与局限于细胞核或细胞体的非特异性DNA链断裂探针不同，这种方法允许在DNA片段化和细胞死亡之前很久检测细胞过程中的细胞骨架片段以及核周体，因此可以作为细胞远端部分（如轴突和树突）中凋亡相关事件的新标记。为了说明这一新工具，我们表明抗体检测阿尔茨海默病中退化神经元和斑块相关小胶质细胞的过程和细胞体。原位检测半胱天冬酶切割的肌动蛋白为评估半胱天冬酶激活在病理和生理过程中的作用提供了一种新的手段。
During apoptosis, activation of a family of cysteine proteases related to interleukin-1beta-converting enzyme (ICE)-related proteases or "caspases" results in endoproteolytic cleavage of multiple substrates at specific aspartate residues. We have sought to develop new antibody probes for the neoepitopes in protein fragments produced by ICE-related proteolytic cleavage as specific markers of events tightly linked to apoptotic mechanisms. Here, we demonstrate that an antibody probe specific for the C terminus of a 32-kd actin fragment produced by ICE-like activity specifically labels apoptotic but not necrotic, differentiated human neuroblastoma cells in culture. Unlike probes for nonspecific DNA strand breaks confined to the nucleus or cell body, this method allows the detection of cytoskeletal fragments in cell processes as well as the perikaryon long before DNA fragmentation and cell death and therefore serves as a novel marker of apoptosis-related events in distal parts of cells such as axons and dendrites. To illustrate this new tool, we show that the antibody detects the processes and cell bodies of degenerating neurons and plaque-associated microglia in Alzheimer's disease. In situ detection of caspase-cleaved actin provides a new means to evaluate the role of caspase activation in pathological and physiological processes.