MEMBRANE PROPERTIES, AMINO ACID RECEPTORS, AND LOCAL CIRCUITS

膜特性、氨基酸受体和局部电路

• 批准号: 3738584
• 负责人: EDWARD F DUDEK
• 金额: --
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3738584
• 关键词: NMDA receptors; action potentials; biological signal transduction; calcium; calcium flux; cerebral cortex; child (0-11); child physical development; child psychology; electroencephalography; electrophysiology; epilepsy; evoked potentials; gamma aminobutyrate; glutamate receptor; hemispherectomy; human tissue; neurons; neuropharmacologic agent; neuropharmacology; neurophysiology; positron emission tomography; pyramidal cells; single cell analysis; sodium; stainings; synapses; voltage /patch clamp

Electrophysiological studies will be conducted on slices from developing human neocortex to examine intrinsic electrophysiological properties of cortical neurons, the pharmacology of excitatory and inhibitory amino acid receptors, and the characteristics of local synaptic circuits. The proposed experiments will be performed on human cortical tissue surgically removed for treatment of catastrophic childhood epilepsy. For tissue from each patient, direct comparisons will be made between the most abnormal area and the least abnormal area. The relative normality of the tissue samples will be evaluated from clinical data, such as intraoperative electrocorticography (ECoG), positron emission tomography (PET), pathology, etc. The experimental methods will include: (1) intracellular recording with current- and voltage-clamp techniques using sharp and patch electrodes, (2) bath and microapplication of pharmacological agonists and antagonists, and (3) intracellular staining. The experiments will test specific hypotheses about the cellular mechanisms of catastrophic childhood epilepsy and about changes in neocortical neurophysiology that occur during human development. Particular emphasis will be on analyses of: (1) current-evoked action potentials and spike bursts and their underlying voltage-dependent Na+ and Ca2+ conductances, (2) N-methyl-D-aspartate (NMDA) receptors, (3) GABA-mediated synaptic inhibition, and (4) local excitatory circuits. Each specific aim includes hypotheses concerning the underlying cellular mechanism(s) of catastrophic childhood epilepsy and developmental changes in the electrophysiology of human cortical neurons. Particular emphasis, especially at the beginning of the grant, will be aimed at testing the hypothesis that epileptogenic and/or developmental changes in human neocortex involve alterations in the number or properties of NMDA receptors or decreases in GABAA-mediated inhibition. Long-term studies will determine possible changes in local excitatory circuits among neocortical pyramidal cells. Alterations in the anatomy of neocortical neurons, which could be associated with epileptogenesis and/or development, will be examined in neurons stained intracellularly with biocytin. The goal is to provide fundamental new information on the electrophysiological characteristics of human cortical neurons, how these properties change during development, and how they are involved in and/or altered during catastrophic childhood epilepsy.

将对发育中的切片进行电生理学研究 人类新皮质检查内在电生理特性 皮质神经元，兴奋性和抑制性氨基酸的药理学 受体和局部突触回路的特征。 这 拟议的实验将通过手术在人体皮质组织上进行 因治疗灾难性儿童癫痫而被移除。 对于来自以下组织的组织 对每个患者最异常的情况进行直接比较 区和异常最少的区域。 组织的相对正常性 样本将根据临床数据进行评估，例如术中 皮质电图 (ECoG)、正电子发射断层扫描 (PET)、病理学、 等。实验方法将包括：（1）细胞内记录 使用锐利和贴片的电流和电压钳技术 电极，（2）药理学激动剂的浴和微量应用以及 拮抗剂，(3)细胞内染色。 实验将测试 关于灾难性童年的细胞机制的具体假设 癫痫以及新皮质神经生理学的变化 人类发展。 特别强调以下方面的分析：(1) 电流诱发的动作电位和尖峰爆发及其潜在的 电压依赖性 Na+ 和 Ca2+ 电导，(2) N-甲基-D-天冬氨酸 (NMDA) 受体，(3) GABA 介导的突触抑制，以及 (4) 局部 兴奋回路。 每个具体目标都包含有关以下方面的假设： 灾难性儿童癫痫的潜在细胞机制和 人类皮质神经元电生理学的发育变化。 特别强调的是，特别是在拨款开始时， 旨在检验致癫痫和/或发育的假设 人类新皮质的变化涉及数量或属性的改变 NMDA 受体的减少或 GABAA 介导的抑制作用的减少。 长期 研究将确定局部兴奋回路可能发生的变化 新皮质锥体细胞。 新皮质解剖结构的改变 神经元，可能与癫痫发生和/或发展有关， 将在细胞内用生物胞素染色的神经元中进行检查。 这 目标是提供有关电生理学的基本新信息 人类皮质神经元的特征，这些特性如何变化 在开发过程中，以及它们如何参与和/或改变 灾难性的儿童癫痫症。