Pilocarpine诱导的小鼠癫痫持续状态导致齿状回未成熟新生神经元形成异常整合  

Immature neurons born one week before pilocarpine-induced status epileptic exhibit aberrant integration at chronic stage after SE

作　　者：袁博 朱坤 胡明 冯改丰 杨蓬勃 张建水 邓美英[3] 刘建新 

机构地区：[1]西安交通大学医学部基础医学院神经生物学研究所,陕西西安710061 [2]西安交通大学医学部基础医学院人体解剖与组织胚胎学系,陕西西安710061 [3]西安交通大学第一附属医院神经内科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2017年第6期785-789,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金面上项目(No.31671249;81171232)~~

摘　　要：目的阐明癫痫持续状态(status epilepticus,SE)发生之前海马产生的未成熟神经元是否在SE之后发生异常整合,包括底树突形成、门区迁移和轴突芽发。方法 Pilocarpine制作小鼠颞叶癫痫模型,在SE诱导前1周采用BrdU或携带绿荧光蛋白的逆转录病毒标记齿状回新生细胞。SE后8周,采用BrdU+Map2和BrdU+NeuN免疫荧光双标染色分别观察分析新生神经元底树突的形成和向齿状回门区的异常迁移。通过齿状回内分子层(inner molecular layer,IML)GFP荧光信号分析观察新生神经元的轴突芽发。采用t-检验分析对比SE组和对照诱导组异常整合细胞的数量。结果对照诱导组动物标记的新生细胞基本未发生异常整合。在SE之后8周,(20.8±8.4)%的新生细胞形成底树突,(15.9±7.4)%的新生细胞发生门区异位迁移,其数量较对照诱导组动物明显增多(P<0.05)。SE组动物齿状回IML出现明显的GFP阳性芽发信号,而在诱导对照组则未出现。结论 SE发生之前海马产生的未成熟神经元在SE之后仍可发生底树突形成、门区异位迁移和轴突芽发等异常整合。Objective To determine whether the integration of immature neurons born before status epilepticus（SE）can be disrupted by an epileptogenic insult.Methods Pilocarpine was used to induce SE in mice.At week 1 before induction,BrdU or retroviral vector expressing green fluorescent protein（RV-GFP）was used to label the newly born cells in the dentate gyrus（DG）.At week 8 after SE,BrdU＋Map2 or BrdU＋NeuN doublelabeling staining was carried out to visualize hilar basal dendrite or hilar ectopic migration.Virus-transduced GFP signals were used to identify the mossy fiber sprouting from the newly generated neurons.The number of cells with aberrant integrations was compared using unpaired Student＇s t-test.Results The percentage of newborn neurons with aberrant dendritic morphology was（20.8±8.4）% at week8 after SE.The percentage of BrdU＋NeuN double labeled cells ectopically migrated into the hilus was（15.9±7.4）%.At week 8 after SE,the chronically epileptic mice showed many GFP＋processes in the IML with the same axonal appearance and small mossy fiber bouton-like structures as those seen in the hilus.The number of newborn neurons with aberrant integrations in SE mice was significantly increased when compared with the control mice（P〈0.05）.Conclusion These data demonstrate the existence of aberrant integrations-hilar basal dendrites,hilar ectopic migration and mossy fiber sprouting in the DGgenerated cells born 1 week before an SE insult.

关 键 词：癫痫持续状态 海马 神经再生 底树突 苔藓纤维芽发 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学]