应用神经逆行示踪及激光显微切割技术精确分离mesostriatal亚群神经元  

Isolation of mesostriatal neurons using neural retrograde tracing and laser-micr-odissection technique

作　　者：杨淑华[1] 高青[1] 李春晓 王超[1] 张静[1] 李贵宝[1] 李振华[1] 孙晋浩[1] 

机构地区：[1]山东大学医学院解剖学教研室,济南250012 [2]山东省潍坊市安丘中医院神经外科,山东潍坊262100

出　　处：《山东大学学报（医学版）》2011年第5期15-18,共4页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金资助项目(30870839);山东省自然科学基金(2R2010HM051)

摘　　要：目的中脑内向纹状体投射的mesostriatal多巴胺能细胞亚群是导致帕金森病的关键神经元,本实验拟寻找该亚群投射神经元的分布及有效分离方法。方法利用立体定位技术将绿色荧光逆行示踪剂注射到新生鼠纹状体内,24h后灌注取脑做冰冻切片;应用免疫荧光组织化学染色方法行酪氨酸羟化酶(TH)染色,采用激光显微切割技术将荧光示踪剂标记且呈TH染色阳性的神经元精确切割并收集。结果荧光示踪的神经元呈TH阳性者即属于mesostriatal亚群神经元,主要分布于中脑黑质,应用激光显微切割技术可精确分离这些逆行示踪且TH染色阳性的神经元。结论应用逆行荧光示踪和激光显微切割技术可精确分离mesostriatal亚群神经元。Objective To explore the distribution of mesostriatal dopaminergic neurons and to establish an effective method to isolate those cells which play key roles in Parkinson′s disease.Methods With stereotaxic apparatus,green retro-beads were accurately injected into the striatum of newborn rats.After 24 h,the brains of the rats were collected and cut into frozen sections after perfusion and fixation.Then dopaminergic neurons were detected by immunohistochemical staining of tyrosine hydroxylase（TH）.And green retro-beads-labeled and TH-positive neurons were isolated by the technique of laser-microdissection.Results Mesostriatal neurons could be labeled with green retro-beads and immunohistochemiscal staining of TH,and were mainly located in the substantia nigra.Moreover,the labeled mesostriatal neurons could be isolated and collected by the technique of laser-micro-dissection.Conclusion Combination of neural retrograde tracing and the laser-micro-dissection technique is an effective and accurate method to isolate mesostriatal dopaminergic neurons.

关 键 词：多巴胺能神经元 神经逆行示踪 激光显微切割技术 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学]