Neuronal stress and injury in C57/BL mice after systemic kainic acid administration
Neuronal stress and injury in C57/BL mice after systemic kainic acid administration
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DOI：
10.1016/s0006-8993(98)00863-4
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发表时间：
1998-11-09
期刊：
影响因子：
2.9
通讯作者：
Cole, AJ
中科院分区：
文献类型：
--
作者：
Hu, RQ;Koh, S;Cole, AJ
红藻氨酸诱导的癫痫作为人类颞叶癫痫的一种模型，由于其行为和病理上的相似性而被广泛研究。虽然红藻氨酸诱导的神经元损伤在大鼠中有很好的特征，但关于红藻氨酸在小鼠中的使用及其后果的数据相对较少。随着转基因小鼠的日益增多，人们开始注意到需要一种具有良好特征的小鼠癫痫发作模型，在该模型中可以检测特定基因操作的效果。因此，我们研究了红藻氨酸在C57/BL小鼠身上的剂量-反应关系和特定组织病理学变化的时间-过程，C57/BL小鼠是转基因技术常用的创立者品系。用红藻氨酸(KA)10~40 mg/kg ip诱导雄性C57/BL小鼠癫痫发作。注射后不同时间点处死动物。使用我们实验室开发的行为量表对癫痫发作进行分级。用原位缺口平移组织化学方法检测DNA片段化，检测神经元损伤程度。在平行实验中，我们使用单抗检测了热休克蛋白家族的可诱导成员HSP-72的表达，HSP-72是神经元损伤的另一个假定标记物。癫痫发作严重程度与海人酸剂量成正比。在较高剂量时，DNA片段化主要出现在CA3区的海马区，24 h内在CA1区、丘脑和杏仁核也有不同程度的DNA片段化，72 h内最大，7 d后基本消失。HSP-72的表达也是高度选择性的，发生在边缘结构中，并且它在一个特有的时间过程中演变。HSP-72主要在DNA断裂的结构中表达。然而，使用双重标记技术，我们发现表达HSP-72和DNA片段的神经元之间基本上没有重叠。这些发现表明DNA片段化和HSP-72的表达是癫痫应激和损伤的补充标志，并支持以下观点：HSP-72的表达在红藻氨酸诱导的癫痫发作后具有神经保护作用。(C)1998 Elsevier Science B.V.保留所有权利。
Kainate-induced seizures are widely studied as a model of human temporal lobe epilepsy due to behavioral and pathological similarities. while kainate-induced neuronal injury is well characterized in rats, relatively little data is available on the use of kainate and its consequences in mice. The growing availability of genetically altered mice has focused attention on the need for well characterized mouse seizure models in which the effects of specific genetic manipulations can be examined. We therefore examined the kainate dose-response relationship and the time-course of specific histopathological changes in C57/BL mice, a commonly used founder strain for transgenic technology. Seizures were induced in male C57/BL mice (kainate 10-40 mg/kg i.p.) and animals were sacrificed at various time-points after injection. Seizures were graded using a behavioral scale developed in our laboratory. Neuronal injury was assayed by examining DNA fragmentation using in situ nick translation histochemistry. In parallel experiments, we examined the expression an inducible member of the heat shock protein family, HSP-72, another putative marker of neuronal injury, using a monoclonal antibody. Seizure severity paralleled kainate dosage. At higher doses DNA fragmentation is seen mainly in hippocampus in area CA3, and variably in CA1, thalamus and amygdala within 24 h, is maximal within 72 h, and is largely gone by 7 days after administration of kainate. HSP-72 expression is also highly selective, occurring in limbic structures, and it evolves over a characteristic time-course. HSP-72 is expressed mainly in structures that also manifest DNA fragmentation. Using double-labeling techniques, however, we find essentially no overlap between neurons expressing HSP-72 and DNA fragmentation. These findings indicate that DNA fragmentation and HSP-72 expression are complementary markers of seizure-induced stress and injury, and support: the notion that HSP-72 expression is neuroprotective following kainate-induced seizures. (C) 1998 Elsevier Science B.V. All rights reserved.