Role of pannexin-1 in the brain and sensory systems

泛连接蛋白1（Panx 1）是泛连接蛋白基因家族的三种膜蛋白之一，其特征在于不同的翻译后蛋白修饰、亚细胞定位和广泛的组织分布，包括脑和感觉器官。目前的知识是Panx 1形成通道，其参与嘌呤能、肾上腺素能、瞬时受体电位香草酸和NMDA受体下游的细胞信号级联。进一步的证据表明，通过升高的细胞外K+和Zn 2+，增加的细胞内Ca 2+，机械刺激，电压，半胱天冬酶介导的裂解，氧化还原调节，激酶，NMDA刺激，或细胞外pH值的变化，Panx 1通道的复杂调节，Panx 1通道代表了一个重要的分子枢纽复杂的调节和多种生理作用，在整个脊椎动物体。** 我的研究计划的长期目标是了解Panx 1在大脑和感觉系统突触传递中的生理作用。目前研究计划的更新有三个目标，测试Panx 1生物学的不同但互补的方面。它们将使用基于细胞的方法进行体外测试，并使用TALEN技术产生的转基因斑马鱼进行体内测试。研究#1探讨了当Panx 1从ER-高尔基体复合体转运到细胞膜时，磷酸化作为“停止-继续”信号的调节作用。当Panx 1到达细胞表面时，目标#2探究细胞外pH如何调节Panx 1通道。目的#3将研究斑马鱼Panx 1基因的遗传切除如何影响多巴胺信号通路，以及为什么这会影响感觉和运动系统。** 总之，我提出了一种高度协同和创新的方法，使用在我的实验室建立的体外和体内方法。预期的结果将提供新的机制洞察Panx 1通道的调节和信号通路的整合汇聚在这个分子枢纽。学员将在各个层面参与这项研究，并在此过程中获得一个充满活力和挑战性的计划广泛的技能。最后，提案中使用的独特斑马鱼模型将提供给其他希望利用我们独特的动物模型进行研究的人，以研究其他基本的生物学问题，或用于毒理学，药物筛选或生物医学/生物工程研究。 **

Pannexin1 (Panx1) is one of three integral membrane proteins of the pannexin gene family characterized by distinct post-translational protein modifications, subcellular localization, and broad tissue distribution including the brain and sensory organs. Current knowledge is that Panx1 forms channels, which are involved in the cell-signaling cascades downstream of the purinergic, adrenergic, transient receptor potential vanilloid, and NMDA receptors. Further evidence suggests a complex regulation of Panx1 channels by elevated extracellular K+ and Zn2+, increased intracellular Ca2+, mechanical stimulation, voltage, caspase-mediated cleavage, redox regulation, kinases, NMDA stimulation, or changes in extracellular pH. Together, Panx1 channels represent a significant molecular hub with complex regulation and multiple physiological roles throughout the vertebrate body. ******The long-term objective of my research program is to understand the physiological role of Panx1 in synaptic transmission in the brain and sensory systems. The renewal of the current research program has three objectives testing distinct, but complementary aspects of Panx1 biology. They will be tested in vitro using cell-based approaches and in vivo using transgenic zebrafish generated by TALEN technology. Objective#1 inquires the regulatory role of phosphorylation as a “stop-go” signal when Panx1 is transported from the ER-Golgi complex to the cell membrane. When Panx1 has arrived at the cell surface, Objective #2 inquires how extracellular pH regulates Panx1 channels. Objective #3 will investigate how genetic ablation of Panx1 genes in the zebrafish affects the dopamine-signaling pathway and why this impacts sensory and motor systems. ******In summary, I propose a highly synergistic and innivative approach using in vitro and in vivo methodology established in my laboratory. The expected outcomes will provide novel mechanistic insight into Panx1 channel regulation and integration of signaling pathways converging on this molecular hub. Trainees will engage in this research at all levels and in the process acquire a broad skill set in a vibrant and challenging program. Finally, the unique zebrafish models used in the proposal will be made available for others who wish to pursue studies exploiting our unique animal models for investigating other fundamental biological questions, or for toxicology, drug screening, or biomedical/bioengineering research. **