Glial ion channels in glia/neurons interactions.

神经胶质/神经元相互作用中的神经胶质离子通道。

• 批准号: 10349558
• 负责人: Laura Bianchi
• 金额: $ 33.58万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10349558
• 关键词: Address; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Behavior; Animal Model; Animals; Antigen-Presenting Cells; Behavior; Behavioral; Behavioral Assay; Bicarbonates; Brain; Buffers; Caenorhabditis elegans; Cell physiology; Chlorides; Data; Demyelinating Diseases; Electrophysiology (science); Elements; Epilepsy; Event; Functional Imaging; Genes; Genetic; Homeostasis; Image; Ion Channel; Ions; Knock-out; Knockout Mice; Lead; Mammals; Mediating; Mediator of activation protein; Methodology; Modeling; Molecular; Multiple Sclerosis; Mutation; Nervous system structure; Neuroglia; Neuronal Dysfunction; Neurons; Nose; Output; Pathology; Permeability; Phenotype; Process; Proteins; Publishing; Regulation; Role; Synapses; System; Testing; Tissues; Touch sensation; Training; Work; behavioral phenotyping; cost; deafness; experimental study; genetic approach; in vivo; interdisciplinary approach; knock-down; knockout animal; mRNA sequencing; mind control; nervous system disorder; novel; reuptake; transcriptome sequencing

Isolated microenvironments like the synapse exist throughout the nervous system where the concentration of ions is regulated by accessory cells, including glia, quite independently from the surrounding tissues. The ionic composition of these microenvironments is key for neuronal function. Despite the fact that glial regulation of ion concentration in microenvironments is a main mode for regulating neuronal activity, our understanding of this type of regulation by glia is limited, especially for ions like Cl- and HCO3-. Furthermore, models are lacking where a comprehensive analysis can be performed on the glial ion channels and transporters involved in regulating ion concentrations, and how these proteins impact neuronal output, from molecule to animal behavior. In our over 10 years of effort aimed at advancing understanding of glia-neurons interaction and its impact on animal behavior using the model C. elegans, we have recently taken the unbiased approach of sequencing the mRNA of Amphid sheath glia. In this application, we propose to establish the mechanism by which one of the identified enriched genes, the glial Cl-/HCO3- permeable channel CLH-1, regulates neuronal output and animal behavior. We previously published that CLH-1 mediates pH regulation in the worm nervous system. Our preliminary results now show that CLH-1 is needed for normal nose-touch behavior. We hypothesize that glial CLH-1 regulates the activity of touch neurons via a direct effect of the permeating ions Cl- and HCO3- on neuronal DEG-1 channels. Thus, the specific aims of this application are: 1) In neurons, to establish the mechanism of neuronal dysfunction when clh-1 is knocked-out, 2) In glia, to determine whether it is the loss of permeation of Cl-, HCO3-, or both that produces the phenotype of clh-1 knock-out worms. Furthermore, in aim 3 we will exploit our proven approach to identify additional glial ion channels and transporter genes that are critical for the glial control of neuronal function and animal behavior: 3) To identify novel glial ion channels and transporters involved in glia-neurons interaction. The importance of regulating ion concentrations in neuronal microenvironments is underscored by the fact that several neurological diseases such as deafness, epilepsy, Alzheimer's, and even demyelinating diseases like multiple sclerosis are characterized by loss of ionic homeostasis. We propose here to use methodologies we have developed and proven to be effective to test mechanisms by which dysregulation of Cl- and HCO3- homeostasis in C. elegans leads to severe neuronal pathology. In addition, we will test the involvement of newly identified genes encoding glial channels and transporters in glia-neurons interaction.

在整个神经系统中都存在孤立的微环境（如突触） 离子受辅助细胞（包括神经胶质）的调节，它与周围的组织完全独立。离子 这些微环境的组成是神经元功能的关键。尽管离子的神经胶质调节 微环境中的浓度是调节神经元活动的主要模式，我们对此的理解 Glia的调节类型受到限制，尤其是对于Cl-和HCO3-等离子。此外，缺乏模型 可以在涉及的神经离子通道和转运蛋白上进行全面分析 调节离子浓度，以及这些蛋白如何影响从分子到动物的神经元输出 行为。在我们十多年的努力中，旨在促进对神经胶质神经元互动及其的理解 使用秀丽隐杆线虫对动物行为的影响，我们最近采取了公正的方法 测序Amphid Sheath Glia的mRNA。在此应用中，我们建议通过 哪一种鉴定出的富集基因，神经胶质Cl-/hCo3-渗透通道Clh-1，调节神经元 产出和动物行为。我们以前发表了CLH-1介导蠕虫神经中的pH调节 系统。现在，我们的初步结果表明，正常的鼻触摸行为需要CLH-1。我们 假设神经胶质CLH-1通过渗透离子Cl-的直接作用调节触摸神经元的活性 和HCO3-在神经元DEG-1通道上。因此，此应用的具体目的是：1）在神经元中， 当CLH-1被敲出时，建立神经元功能障碍的机制，2）在Glia中，以确定它是否是否 是Cl-，HCO3-的渗透的丧失，或两者都会产生CLH-1敲除蠕虫的表型。 此外，在AIM 3中，我们将利用我们验证的方法来识别其他神经离子渠道和 转运蛋白基因对神经元功能和动物行为的神经胶质控制至关重要：3） 与神经胶质神经元相互作用有关的新型神经离子通道和转运蛋白。调节离子的重要性 神经元微环境中的浓度强调了几种神经系统疾病 例如耳聋，癫痫，阿尔茨海默氏症甚至脱髓鞘性疾病，例如多发性硬化症 以离子稳态的流失为特征。我们在这里建议使用我们开发的方法论，并 被证明对测试机制有效 导致严重的神经元病理。此外，我们将测试新鉴定的编码基因的参与 神经神经元相互作用中的神经胶质通道和转运蛋白。