Induction of Temporal Lobe Epilepsy in Mice with Pilocarpine
Induction of Temporal Lobe Epilepsy in Mice with Pilocarpine
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DOI：
10.21769/bioprotoc.3533
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发表时间：
2020-02-20
期刊：
影响因子：
0.8
通讯作者：
Naegele, Janice R.
中科院分区：
文献类型：
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作者：
Arshad, Muhammad N.;Naegele, Janice R.
在啮齿类动物颞叶癫痫（TLE）的匹罗卡品模型中，全身注射匹罗卡品会引起持续、长时间的边缘癫痫发作，这种情况被称为“癫痫持续状态”（SE）。在适当剂量下，许多近交系小鼠在注射匹罗卡品后一小时内表现出行为癫痫。使用基于原始拉辛量表修改的行为评分系统，人们可以监测癫痫发作的行为，因为它们发展为更长的癫痫发作和SE。SE通常与海马神经元亚群的损伤和海马的其他结构变化有关，通常会自行消退。然而，更精确地控制SE持续时间通常是通过在SE发作后1至3小时向小鼠注射苯二氮卓类药物来抑制癫痫发作。在匹罗卡品诱发SE后几天，电图和行为癫痫开始自发发生。该方案的目标是可靠地产生发生自发性复发性癫痫发作（SRS）的小鼠，并显示出严重人类内侧颞叶癫痫（mTLE）的典型脑神经病理变化，没有高死亡率。为了降低死亡率，多次阈下注射匹罗卡品，这增加了小鼠发生SE的百分比，但没有伴随死亡。通过使用苯二氮卓类药物咪达唑仑（Versed）抑制SE，可以精确控制SE持续时间（1或3小时）。我们发现，这种方法是一种有效的方法，可以培养出具有人类mTLE特征的小鼠，包括高频间歇峰和波活动以及SRS。此外，我们和其他人已经证明，该方案产生小鼠海马gaba能中间神经元亚群的兴奋性毒性细胞死亡，特别是在齿状回和齿状颗粒细胞兴奋性突起的代偿性发芽（苔藓纤维发芽）。该协议的各个方面已在我们之前的几篇出版物中进行了描述。
In the pilocarpine model of temporal lobe epilepsy (TLE) in rodents, systemic injections of pilocarpine induce continuous, prolonged limbic seizures, a condition termed "Status Epilepticus" (SE). With appropriate doses, many inbred strains of mice show behavioral seizures within an hour after pilocarpine is injected. With the behavioral scoring system based on a modification of the original Racine scale, one can monitor the seizures behaviorally, as they develop into more prolonged seizures and SE. SE is typically associated with damage to subsets of hippocampal neurons and other structural changes in the hippocampus and generally subsides on its own. However, more precise control of the duration of SE is commonly achieved by injecting a benzodiazepine into the mouse 1 to 3 h after the onset of SE to suppress the seizures. Several days following pilocarpine-induced SE, electrographic and behavioral seizures begin to occur spontaneously. The goal of this protocol is to reliably generate mice that develop spontaneous recurrent seizures (SRS) and show the typical neuropathological changes in the brain characteristic of severe human mesial temporal lobe epilepsy (mTLE), without high mortality. To reduce mortality, multiple subthreshold injections of pilocarpine are administered, which increases the percentage of mice developing SE without concomitant mortality. Precise control of the duration of SE (1 or 3 h) is achieved by suppressing SE with the benzodiazepine Midazolam (Versed). We have found that this protocol is an efficient means for generating mice that subsequently develop characteristics of human mTLE including high -frequency interictal spike and wave activity and SRS. In addition, we and others have shown that this protocol produces mice that show excitotoxic cell death of subsets of hippocampal GABAergic interneurons, particularly in the dentate gyrus and compensatory sprouting of excitatory projections from dentate granule cells (mossy fiber sprouting). Aspects of this protocol have been described in several of our previous publications.