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山东大学学报(医学版) ›› 2012, Vol. 50 ›› Issue (7): 14-18.

• 基础医学 • 上一篇    下一篇

中脑皮层投射多巴胺神经元的提取及帕潘立酮对其电生理学特性的影响

董海曼1,高青1,张静1,暴丽华1,李贵宝1,彭雷1,冯晓雯1,
岳庆伟1,刘增训2,孙晋浩1,高英茂1   

  1. 1.山东大学医学院人体解剖与组织胚胎学研究所, 济南 250012;
    2.山东省精神卫生中心精神科, 济南 250001
  • 收稿日期:2012-02-13 出版日期:2012-07-10 发布日期:2012-07-10
  • 通讯作者: 孙晋浩(1973- ),男,教授,博士生导师,主要从事神经发育与神经再生的研究。 E-mail:sunjinhao@gmail.com
  • 作者简介:董海曼(1987- ),女,硕士研究生,主要从事神经发育与神经再生的研究。 E-mail:haiman19879@126.com
  • 基金资助:

    国家自然科学基金(81071081);山东省自然科学基金(ZR2010HM051);山东省科技发展计划(2011GSF11810)

Extraction of mesocortical dopaminergic projection neurons and effects of paliperidone on their electrophysiological properties

DONG Hai-man1, GAO Qing1, ZHANG Jing1, BAO Lihua1, LI Gui-bao1, PENG Lei1,
FENG Xiao-wen1, YUE Qing-wei1, LIU Zeng-xun2, SUN Jin-hao1, GAO Ying-mao1   

  1. 1. Institute of Human Anatomy and Embryology, School of Medicine, Shandong University, Jinan 250012, China;
    2. Department of Psychiatry, Shandong Mental Health Center, Jinan 250001, China
  • Received:2012-02-13 Online:2012-07-10 Published:2012-07-10

摘要:

目的   精确示踪并分离中脑皮层多巴胺(DA)能神经元,检测帕潘立酮对其动作电位的影响。方法   采用脑立体定位技术,将荧光逆行性示踪剂注射至鼠脑内嗅皮层区,灌注取脑,行冰冻切片;采用免疫组织化学技术进行酪氨酸羟化酶(TH)染色;依据荧光强度分选示踪标记的神经细胞并培养,采用细胞膜片钳技术对其进行电流注射,并加入抗精神病药物帕潘立酮,检测加药前后动作电位的产生及变化。结果   准确标记向脑皮质投射的中脑DA能神经元,呈TH阳性,位于腹侧被盖区;筛选出标记的DA能神经元,电流刺激可诱发出DA神经元典型的动作电位,加入抗精神病药物帕潘立酮后,动作电位幅度降低(P<0.05)、宽度变小(P<0.01)。结论   精确分离出向脑额叶投射的中脑皮层DA神经元,帕潘立酮可降低此类细胞动作电位幅度及宽度,从而降低细胞的兴奋性。

关键词: 2型糖尿病;筛查;空腹毛细血管血糖;空腹血浆血糖;糖负荷2h血浆血糖;糖化血红蛋白;危险因素评分表;受试者工作特征, 帕潘立酮;多巴胺;神经元;立体定位技术;动作电位

Abstract:

Objective   To precisely trace and separate mesocortical dopaminergic (DA) neurons and detect the effects of paliperidone on their action potentials. Methods   According to stereotaxic apparatus, fluorescent retrograde tracer was injected to the entorhinal cortex of rats′ brains. The brains were removed after being perfused to be made into frozen sections. Immunohistochemistry was adopted for tyrosine hydroxylase (TH) staining. The nerve cells labeled by the tracer were sorted by the fluorescence intensity, and cultured. The generations and changes of action potentials before and after antipsychotic paliperidone treatment were detected by using cell patch-clamp technique for the current injection. Results   The midbrain DA neurons which projected to the cerebral cortex were labeled accurately, showing TH-positive, and located in the ventral tegmental area. DA neurons which were marked and filtered out showed a typital action potential after being induced by electrical stimulation. After antipsychotic paliperidone treatment, the amplitude of action potential appeared lower(P<0.05), and the width became smaller(P<0.01). Conlusion   The mesocortical DA neurons which project to the frontal brain can be separate accurately. Paliperidone can reduce the amplitude and width of the action potentials of the cells, thus reduce the excitability of the cells.

Key words: Type 2 diabetes mellitus; Screening; Fasting capillary blood glucose; Fasting plasma glucose; 2-hour plasma glucose; Glycosylated haemoglobin; Diabetes risk score; Receiver operating characteristic, Paliperidone; Dopamine; Neurons; Stereotaxic techniques; Action potentiais

中图分类号: 

  • R338.2
[1] 王孝勇1,李士雪2,孙秀彬2,逄增昌3,乔青4. 5种方法筛查无症状2型糖尿病的效果评价[J]. 山东大学学报(医学版), 2011, 49(4): 158-162.
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