SYNCHRONIZED RHYTHMIC ACTIVITY OF THE PALLIDUM

苍白球的同步节律活动

• 批准号: 6595224
• 负责人: Hitoshi Kita
• 金额: $ 11.11万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6595224
• 关键词: Parkinson's disease; brain electrical activity; calcium binding protein; calcium channel; calcium indicator; digital imaging; disease /disorder model; dopamine; electron microscopy; electrostimulus; immunocytochemistry; laboratory rat; lenticular nucleus; membrane potentials; microelectrodes; neural conduction; neural transmission; neuroanatomy; neuropharmacology; potassium channel; subthalamus; synapses; tissue /cell culture; voltage /patch clamp; voltage gated channel

A loss of midbrain dopaminergic cells results in parkinsonism. It has been hypothesized that parkinsonian rigidity and tremor are associated with abnormal, wide spread synchronized, rhythmic firing of the pallidum. The goal of this proposal is to reveal the mechanisms responsible for the synchronized rhythmic activity of the pallidum. Our basic hypothesis is that the synchronized rhythmic activity is generated because dopamine (DA) depletion has altered the intrinsic membrane properties and synaptic inputs of the pallidum. We hypothesize that pallidal neurons have intrinsic mechanisms for membrane oscillation, and that the membrane oscillatory mechanism is controlled by the DAergic innervation from the midbrain and peptidergic innervations from the neostriatum (Str). To test these hypothesis, physiological and pharmacological studies will be performed using a whole cell recording method in slice preparations. Our study will focus on the potassium and calcium currents involved in the membrane oscillations and effects of DNA and striatal peptides on these currents. Calcium imaging will aid in analyzing the distribution of calcium currents along the dendrites. We hypothesize that the wide synchronization of activity seen in the pallidum of parkinsonian subjects is due to changes in synaptic transmission. Evaluation of this hypothesis should begin with physiological and anatomical studies. To reveal the properties of unitary post-synaptic responses of pallidal neurons to striatal, intra-pallidal, and subthalamic stimulation, and to reveal the effects of DA and striatal peptides on the strength and duration of unitary post-synaptic responses, physiological and pharmacological studies will be performed using a whole cells recording method in slice preparations. All neurons record in the slice experiments will be further characterized for parvalbumin immunoreactivity, somato-dendritic morphology, and at the molecular level for ion channels, receptors, and calcium binding proteins using single cell RT-PCR techniques. To reveal the structural bases for the pallidal synchronizations, intracellular staining of striatal, pallidal, and subthalamic neurons will be performed in anesthetized rats, and the axons of stained neurons will then be analyzed, using computer assisted 3-dimensional reconstruction systems, at both the light and electron microscopic levels. The light microscopic analysis will reveal the number and distribution of synaptic boutons of single axons in the pallidum, and the electron microscopic analysis will reveal the ultrastructure of the synapses, the synaptic pattern, and the structure post-synaptic to the synapses.

中脑多巴胺能细胞的丧失会导致帕金森氏症。一直以来 假设帕金森强直和震颤与 苍白球异常、大范围、同步、有节奏的放电。这个 这项提案的目标是揭示导致 苍白球的同步节律活动。我们的基本假设是 同步节律活动的产生是因为多巴胺(DA) 耗竭改变了固有的膜特性和突触 苍白球的输入。 我们假设苍白球神经元有内在的机制 膜振荡，膜振荡机制是 受中脑和多肽能神经支配 新纹状体的神经支配(Str)。为了检验这些假设， 生理和药理学研究将使用一个完整的 切片制备中的细胞记录方法。我们的研究将集中在 钾和钙电流参与膜振荡和 DNA和纹状体多肽对这些电流的影响。钙质成像 将有助于分析钙电流沿线的分布。 树枝状结构。 我们假设，大范围的同步性活动 帕金森病患者的苍白球是由于突触的变化 变速箱。对这一假设的评估应该从 生理和解剖学研究。揭示酉性的性质 苍白球神经元对纹状体、苍白球内、 和丘脑底核刺激，揭示多巴胺和纹状体的作用 对一元性突触后反应强度和持续时间的影响， 生理和药理学研究将使用一个完整的 切片制备中的细胞记录方法。所有神经元都记录在 切片实验将进一步表征小白蛋白。 免疫反应性、体树突状细胞形态和分子水平 用于离子通道、受体和钙结合蛋白 细胞RT-PCR技术。 为了揭示苍白球同步的结构基础， 纹状体、苍白球和丘脑下部神经元的细胞内染色将 在麻醉的大鼠身上进行，染色神经元的轴突将 然后进行分析，采用计算机辅助的三维重建 系统，在光和电子显微镜水平上。那盏灯 显微分析将揭示突触的数量和分布 苍白球内单个轴突的突起和电子显微镜 分析将揭示突触的超微结构，突触 模式，以及突触后的结构与突触。