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。
Objective To precisely trace and separate mesocortical dopaminergic (DA) neurons and detect the effects of palipeddone 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 fro- zen 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 be- fore and after antipsychotic paliperidone treatment were detected by using cell patch-clamp technique for the current in- jection. Results The midbraln 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 andwidth of the action potentials of the cells, thus reduce the excitability of the cells.
出处
《山东大学学报(医学版)》
CAS
北大核心
2012年第7期14-18,共5页
Journal of Shandong University:Health Sciences
基金
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)