Kainic acid-induced seizures produce necrotic, not apoptotic, neurons with internucleosomal DNA cleavage: Implications for programmed cell death mechanisms
Kainic acid-induced seizures produce necrotic, not apoptotic, neurons with internucleosomal DNA cleavage: Implications for programmed cell death mechanisms
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DOI：
10.1016/s0306-4522(00)00085-3
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发表时间：
2000-01-01
期刊：
影响因子：
3.3
通讯作者：
Cai, B
中科院分区：
文献类型：
--
作者：
Fujikawa, DG;Shinmei, SS;Cai, B
由红藻氨酸诱导的长时间癫痫发作（癫痫持续状态）激活程序性细胞死亡机制，并且认为红藻氨酸诱导的癫痫持续状态诱导神经元凋亡。为了验证这一假设，成年大鼠进行3小时红藻氨酸诱导的癫痫发作，与24或72小时的恢复期。通过苏木精-伊红染色和原位末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL染色）的光学显微镜，电子显微镜，以及从五个脆弱的大脑区域提取的DNA琼脂糖凝胶电泳来评估神经元死亡。自发和MK-801诱导的凋亡神经元从新生大鼠的压后皮质，通过光学和电子显微镜评价，作为阳性对照细胞凋亡。令人惊讶的是，凋亡神经元的大染色质团块是TUNEL阴性的，而细胞质显示轻至中度TUNEL染色，与超微结构上缺乏可识别的核膜以及细胞核和细胞质内容物的混合一致。超微结构上，由海人酸诱导的癫痫持续状态产生的嗜酸性神经元，经苏木精-伊红染色鉴定，为深色、萎缩和坏死，核固缩，含有小的分散的染色质团块，胞质空泡，其中一些肿胀，线粒体破坏。未见凋亡细胞。在检查的23个大脑区域中，多达20个区域发现了嗜酸神经元，占检查神经元总数的10-25%。这些神经元的子集（
Prolonged seizures (status epilepticus) induced by kainic acid activate programmed cell death mechanisms, and it is believed that kainic acid-induced status epilepticus induces neuronal apoptosis. In order to test this hypothesis, adult rats were subjected to 3-h kainic acid-induced seizures, with 24- or 72-h recovery periods. Neuronal death was assessed by light microscopy with the Hematoxylin and Eosin stain and with in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL stain), by electron microscopy, and by agarose gel electrophoresis of DNA extracted from five vulnerable brain regions. Spontaneous and MK-801-induced apoptotic neurons from retrosplenial cortex of neonatal rats, evaluated by light and electron microscopy, were used as positive controls for apoptosis. Surprisingly, the large chromatin clumps of apoptotic neurons were TUNEL negative, whereas the cytoplasm showed light-to-moderate TUNEL staining, consistent with a lack of identifiable nuclear membranes ultrastructurally, and with intermingling of nuclear and cytoplasmic contents. Ultrastructurally, the acidophilic neurons produced by kainic acid-induced status epilepticus, identified with Hematoxylin and Eosin stain, were dark, shrunken and necrotic, with pyknotic nuclei containing small, dispersed chromatin clumps, and with cytoplasmic vacuoles, some of which were swollen, disrupted mitochondria. No apoptotic cells were seen. Acidophilic neurons were found in up to 20 of 23 brain regions examined and comprised 10-25% of the total number of neurons examined. A subset of these neurons (