Glial ion channels in glia/neurons interactions.

像突触这样的孤立微环境存在于整个神经系统中， 离子的调节是由辅助细胞，包括神经胶质细胞，完全独立于周围组织。离子 这些微环境的组成是神经元功能的关键。尽管神经胶质细胞对离子的调节 微环境中的浓度是调节神经元活动的主要模式，我们对此的理解是， 神经胶质细胞的调节类型是有限的，特别是对于Cl-和HCO 3-等离子。此外，缺乏模型 其中可以对参与的神经胶质离子通道和转运蛋白进行综合分析， 调节离子浓度，以及这些蛋白质如何影响神经元输出，从分子到动物 行为在我们超过10年的努力，旨在促进了解胶质神经元的相互作用及其 使用模型C对动物行为的影响。elegans，我们最近采取了公正的方法， Amphid鞘胶质细胞的mRNA测序。在本申请中，我们建议通过以下方式建立该机制： 其中一个被鉴定的富集基因，神经胶质Cl-/HCO 3-渗透通道CLH-1，调节神经元 产量和动物行为。我们以前发表过CLH-1介导蠕虫神经系统中的pH调节， 系统我们的初步结果表明，CLH-1是正常的鼻子触摸行为所必需的。我们 假设胶质细胞CLH-1通过渗透离子Cl-的直接作用调节触觉神经元的活性， 和HCO 3-在神经元DEG-1通道上。因此，本申请的具体目的是：1）在神经元中， 建立clh-1敲除后神经元功能障碍的机制，2）在胶质细胞中，以确定其是否 是Cl-、HCO 3-或两者的渗透损失，其产生clh-1敲除蠕虫的表型。 此外，在目标3中，我们将利用我们已证实的方法来识别其他神经胶质细胞离子通道， 对神经元功能和动物行为的神经胶质控制至关重要的转运蛋白基因：3）鉴定 参与神经胶质-神经元相互作用的新型神经胶质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被几种神经系统疾病 如耳聋、癫痫、阿尔茨海默氏症，甚至脱髓鞘疾病如多发性硬化症， 以失去离子平衡为特征。我们在这里建议使用我们开发的方法， 被证明是有效的测试机制，其中失调的Cl-和HCO 3-稳态在C。elegans 导致严重的神经元病变此外，我们还将检测新发现的编码 神经胶质-神经元相互作用中的神经胶质通道和转运蛋白。

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.