Cloning and expression of sodium channel beta subunits

钠通道β亚基的克隆和表达

• 批准号: 6921979
• 负责人: AMANDA J. HOBSON
• 金额: $ 3.34万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6921979
• 关键词: alternatives to animals in research; ankyrins; antisense nucleic acid; biological signal transduction; cell adhesion molecules; cytoskeleton; electrophysiology; gene expression; genetic models; genetically modified animals; immunocytochemistry; intracellular transport; model design /development; molecular cloning; myelinopathy; neural transmission; neurogenesis; node of Ranvier; predoctoral investigator; protein localization; protein structure function; protein transport; sodium channel; voltage gated channel; zebrafish

DESCRIPTION (provided by applicant): The goal of this research project is to test the hypothesis that voltage gated sodium channel beta subunits are cell adhesion molecules in vivo, and are involved in the formation of nodes of Ranvier as well as the recruitment of other molecules involved in the nodal signaling complex. An alternative model system using Zebrafish is being generated in our laboratory. First, the sodium channel beta subunits in Zebrafish will be cloned and characterized using an in vitro system and electrophysiological techniques. Preliminary data show that Zebrafish have beta subunits orthologous to higher vertebrates as well as novel subunits not previously characterized. Next, "knock down" fish will be generated which will allow us to examine the role of beta subunits in vivo using antisense RNA techniques. Finally, a dominant negative beta1subunit will be used to test the ability of the beta1subunit to recruit ankyrin to the plasma membrane. Understanding the mechanism of node of Ranvier formation is important to elucidating the mechanisms of demyelinating disease. Demyelinating diseases are characterized by a loss of myelin and an impairment of action potential conduction as nodes of Ranvier are lost. Understanding nodal formation can contribute to clarifying the triggers for demyelinating disease as well as potential therapies.

描述（由申请人提供）：该研究项目的目的是测试以下假设：电压门通道元素β亚基是体内的细胞粘附分子，并且参与了Ranvier的节点以及其他分子的募集。我们的实验室正在生成使用斑马鱼的替代模型系统。首先，使用体外系统和电生理技术将斑马鱼中的钠通道β亚基克隆和表征。初步数据表明，斑马鱼与较高脊椎动物的β亚基以及以前未表征的新型亚基。接下来，将生成“敲除”鱼，这将使我们能够使用反义RNA技术在体内检查β亚基的作用。最后，将使用主要的负beta1subunit来测试beta1subunit募集Ankyrin到质膜的能力。了解兰维尔形成节点的机制对于阐明脱髓鞘疾病的机制很重要。脱髓鞘性疾病的特征是髓磷脂的丧失，并且随着兰维尔节点的流失，动作潜在传导的损害。了解淋巴结的形成可以有助于阐明脱髓鞘疾病以及潜在疗法的触发因素。