TRPV4: Mechanosensitivity and Skeletal Dysplasia

TRPV4：机械敏感性和骨骼发育不良

• 批准号: 8023251
• 负责人: CHING KUNG
• 金额: $ 28.19万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8023251
• 关键词: Alleles; Amino Acids; Animals; Astronauts; Beds; Blood Pressure; Bone Development; Charcot-Marie-Tooth Disease; Chimera organism; Clinical; Cultured Cells; Defect; Disease; Dissection; Documentation; Dysplasia; Electrophysiology (science); G Protein-Coupled Receptor Genes; Genetic; Growth; Health; Hearing; Human; Ion Channel; Knowledge; Life; Lipids; Literature; Mechanics; Membrane; Modeling; Molecular; Molecular Models; Mus; Mutate; Mutation; Oocytes; Osmolar Concentration; Osteoporosis; Pathology; Patients; Peptides; Phenotype; Physiology; Poisoning; Proteins; Rattus; Simulate; Smell Perception; Spinal Muscular Atrophy; Stretching; Structure; TRP channel; Taste Perception; Techniques; Testing; Time; Touch sensation; Vision; Weight; Work; Xenopus oocyte; Yeasts; base; bone mass; design; expectation; gain of function; gain of function mutation; human disease; improved; interdisciplinary approach; molecular modeling; mutant; patch clamp; response; skeletal dysplasia; theories; voltage; yeast genetics

Unlike vision, smell, and tastes, the molecular mechanisms of hearing, touch, and blood- pressure or systemic-osmolarity sensing are unknown. How ion channels receive mechanical forces remain obscure, except for bacterial channels MscL, MscS. We are using the rat TRP channel V4 subtype, to help fill this knowledge gap. We have now shown TRPV4's direct response to membrane stretch under Xenopus oocyte patch clamp. We have also developed yeast as a new arena to dissect TRPV4. Our work challenges a previous TRPV4's osmotic-gating model. We plan to define the force-sensing domain within TRPV4, using strategic peptide insertions and chimeras. 2008-11 found TRPV4 gain-of-function mutations to cause two sets of human diseases: skeletal dysplasia (SD) and adult peripheral neuropathy. We examined 14 SD alleles and found the level of increased basal TRPV4 activity parallels the severity of the SD disease. Besides understanding the working of the wild-type TRPV4 channels, we now plan to examine 17 mutations in the ankyrin-repeat domain (ARD), of which crystal structure is known. This work aims towards understanding how ARD mutations can lead to two disparate diseases. In collaboration, we will also use trpV4-/- knock-out mice to test the role of TRPV4 in fracture healing.

与视觉、嗅觉和味觉不同，听觉、触觉和血液的分子机制- 压力或全身渗透压传感是未知的。离子通道如何接收机械信号 力仍然模糊，除了细菌通道MscL，MscS。我们用老鼠的TRP channel V4 subtype，to help fill填补this knowledge知识gap差距.我们现在已经证明了TRPV 4的直接 在爪蟾卵母细胞膜片钳下对膜拉伸的反应。我们还开发了 酵母作为一个新的竞技场来剖析TRPV 4。我们的工作挑战了以前的TRPV 4的自动门控 模型我们计划使用战略肽来定义TRPV 4中的力感应结构域， 插入和嵌合体。 2008-11发现TRPV 4功能获得性突变导致两组人类疾病： 骨骼发育不良（SD）和成人周围神经病变。我们检测了14个SD等位基因，发现 基础TRPV 4活性的增加水平与SD疾病的严重程度相平行。除了 了解野生型TRPV 4通道的工作，我们现在计划检查17 锚定重复结构域（ARD）中的突变，其晶体结构已知。这项工作 旨在了解ARD突变如何导致两种不同的疾病。在 合作，我们还将使用trpV 4-/-敲除小鼠来测试TRPV 4在骨折中的作用 治愈