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

山东大学学报(医学版) ›› 2011, Vol. 49 ›› Issue (5): 15-18.

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

杨淑华1，高青1，李春晓2，王超1，张静1，李贵宝1，李振华1，孙晋浩1

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

收稿日期:2010-10-12 出版日期:2011-05-10 发布日期:2011-05-10
通讯作者: 孙晋浩（1970- ），男，教授，主要从事神经发育与神经再生方面的研究。 E-mail:sunjinhao＠sdu.edu.cn
作者简介:杨淑华（1986- ），女，硕士研究生，主要从事神经发育与神经再生的研究。
基金资助:
国家自然科学基金资助项目（30870839）；山东省自然科学基金（2R2010HM051）

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

YANG Shu-hua1, GAO Qing1, LI Chun-xiao2, WANG Chao1, ZHANG Jing1, LI Gui-bao1, LI Zhen-hua1, SUN Jin-hao1

1. Department of Anatomy, School of Medicine， Shandong University， Jinan 250012， China;
2. Department of Neurosurgery, Anqiu Hospital of Traditional Chinese Medicine, Weifang 262100, Shandong, China

Received:2010-10-12 Online:2011-05-10 Published:2011-05-10

摘要/Abstract

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。

关键词: 多巴胺能神经元; 神经逆行示踪; 激光显微切割技术

Abstract:

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 24h, 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. ResultsMesostriatal 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.

Key words: Dopaminergic neurons; Neural retrograde tracing; Laser-microdissection technique

中图分类号:

R322.8

杨淑华1，高青1，李春晓2，王超1，张静1，李贵宝1，李振华1，孙晋浩1. 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[J]. 山东大学学报(医学版), 2011, 49(5): 15-18.

YANG Shu-hua1, GAO Qing1, LI Chun-xiao2, WANG Chao1, ZHANG Jing1, LI Gui-bao1, LI Zhen-hua1, SUN Jin-hao1. Isolation of mesostriatal neurons using neural retrograde tracing and laser-micr-odissection technique[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2011, 49(5): 15-18.

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