EXPERIMENTAL STUDIES IN SENSORY PATHWAYS

感觉通路的实验研究

• 批准号: 2609582
• 负责人: STEPHEN T KITAI
• 金额: $ 23.61万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-08-01 至 2001-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2609582
• 关键词: acetylcholine; action potentials; basal ganglia; biocytin; brain mapping; cell membrane; central neural pathway /tract; dopamine; electron microscopy; electrophysiology; fluorescent dye /probe; glutamates; histochemistry /cytochemistry; immunocytochemistry; laboratory rat; neuroanatomy; neurophysiology; sensory mechanism; single cell analysis; substantia nigra; tissue /cell culture; voltage /patch clamp

The proposed work aims to extend the high resolution of single neuron analysis with combined anatomical and immunocytochemical techniques for a more precise description of cellular, electrical membrane and ionic currents; neurotransmitter and neuromodulator action; and circuits of neuronal elements in the basal ganglia. Moreover, at the same time, the proposed work aims to provide a more meaningful interpretation of various nuclear functions of the basal ganglia than can be obtained by conventional single or multiple unit recording or by conventional anatomical technique. The areas of interest will be addressed with intensive usage of the well established methodology in our laboratory, whereby, intracellular recording and intracellular labeling with biocytin and other labeling materials are combined with subsequent light and electron microscopic analysis. In addition, histochemical methods, including retrograde and anterograde transport techniques, will be employed. This convergent multidisciplinary analysis will be directed to in vitro slice preparation and cultured and acutely dissociated neuron preparation obtained from rat neostriatum globus pallidus, entopeduncular nucleus, substantia nigra, and subthalamus to three sets of aims: The first set will characterize the membrane properties (including ionic conductances such as K, Na, Ca, etc.) of the neuron by the use of whole-cell (voltage) clamp technique in acutely dissociated or cultured neurons or by an intracellular current clamp recording in slice preparation. The second set will delineate the action of putative neurotransmitters or neuromodulators by monitoring changes in ionic conductances produced by the drugs or their agonists and antagonists. The transmitters to be studied are dopamine (DA), glutamate, and acetylcholine (ACh), which are the major known neurotransmitters and modulators involved in these nuclear operation. We will identify the transmitter and morphological phenotypes of the neurons under study by anatomical techniques by combined techniques of immunocytochemistry. The third set is to re-examine functional circuits involved among these nuclei (e.g., subthalamo-pallidal, striato-nigral, etc.) by trigger-averaging techniques with simultaneous recordings from the projection and its target neurons. Disequilibria of cholinergic, dopaminergic, glutaminergic, and GABAergic systems in the basal ganglia are believed involved in the pathophysiology of basal ganglia disease. The search for more specific treatments will be facilitated by a clearer understanding of the specific nature of the basal ganglia anatomy, physiology, pharmacology, and subsequent imbalance of extra-pyramidal operation.

拟议的工作旨在扩展单个神经元的高分辨率 结合解剖学和免疫细胞化学技术进行分析 更精确地描述细胞、电膜和离子 电流；神经递质和神经调节剂的作用；和电路 基底神经节中的神经元元件。 而且，与此同时， 拟议的工作旨在为各种问题提供更有意义的解释 基底神经节的核功能比常规方法所能获得的 单个或多个单元记录或通过传统的解剖技术。 将通过密集使用该井来解决感兴趣的领域 我们的实验室建立了方法论，据此，细胞内记录 以及用生物胞素和其他标记材料进行细胞内标记 结合随后的光学和电子显微镜分析。 在 此外，组织化学方法，包括逆行和顺行 将采用运输技术。 这种融合的多学科分析将针对体外 切片制备以及培养和急性分离神经元制备 从大鼠新纹状体苍白球、内脚核获得， 黑质和底丘脑实现三组目标： 第一组将表征膜特性（包括离子 神经元的电导（例如 K、Na、Ca 等） 急性解离或培养中的全细胞（电压）钳技术 神经元或通过切片中的细胞内电流钳记录 准备。 第二组将描述假定的神经递质或 通过监测神经调节剂产生的离子电导的变化 药物或其激动剂和拮抗剂。 待研究的发射机 多巴胺（DA）、谷氨酸和乙酰胆碱（ACh）是主要的 已知的神经递质和调节剂参与这些核操作。 我们将鉴定递质和形态表型 通过解剖技术通过组合技术研究的神经元 免疫细胞化学。 第三组是重新检查这些之间涉及的功能电路 核（例如，丘脑-苍白球核、纹状体-黑质核等） 触发平均技术与同时记录 投射及其目标神经元。 胆碱能、多巴胺能、谷氨酰胺能和 GABA 能的不平衡 基底神经节中的系统被认为参与病理生理学 基底神经节疾病。 寻找更具体的治疗方法将是 有助于更清楚地了解基础的具体性质 神经节解剖学、生理学、药理学以及随后的不平衡 锥体外系手术。