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，在那里集中的 离子由包括神经胶质细胞在内的辅助细胞独立于周围组织进行调节。《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方式，我们对此的理解 胶质细胞的调节类型是有限的，特别是对Cl-和HCO3-这样的离子。此外，还缺乏模型。 在那里可以对参与的神经胶质细胞离子通道和转运蛋白进行全面的分析 调节离子浓度，以及这些蛋白质如何影响从分子到动物的神经元输出 行为。在我们10多年的努力中，我们致力于促进对神经胶质细胞-神经元相互作用及其 使用线虫模型对动物行为的影响，我们最近采取了公正的方法 对两性鞘神经胶质细胞的mRNA进行测序。在本申请中，我们建议通过以下方式建立该机制 已鉴定的丰富基因中的哪一个，胶质细胞氯/HCO3通透性通道CLH-1，调节神经元 产量和动物行为。我们之前发表了CLH-1在蠕虫神经中介导pH调节的文章 系统。我们的初步结果表明，正常的鼻子触摸行为需要CLH-1。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动 和神经元DEG-1通道上的HCO3-。因此，这一应用的具体目标是：1)在神经元中， 建立CLH-1被敲除时神经元功能障碍的机制，2)在胶质细胞中，以确定是否 是氯离子和/或HCO3-的渗透性丧失，从而产生CLH-1敲除蠕虫的表型。 此外，在目标3中，我们将利用我们成熟的方法来识别更多的胶质细胞离子通道和 神经胶质控制神经元功能和动物行为的关键转运蛋白基因：3)鉴定 参与神经胶质细胞-神经元相互作用的新型神经胶质离子通道和转运体。规范离子的重要性 神经微环境中的浓度被以下事实所强调：几种神经系统疾病 如耳聋、癫痫、阿尔茨海默氏症，甚至多发性硬化症等脱髓鞘疾病 以离子动态平衡丧失为特征的。我们在这里建议使用我们开发的方法和 证明能有效地检测线虫体内氯离子和HCO3-动态平衡失调的机制 会导致严重的神经元病变。此外，我们将测试新发现的编码基因的参与程度 神经胶质细胞-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.