Molecular and Genetic Analysis of Fin Regeneration in Zebrafish

斑马鱼鳍再生的分子和遗传分析

• 批准号: 10204054
• 负责人: Junsu Kang
• 金额: $ 37.71万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10204054
• 关键词: Address; Affect; Afferent Neurons; Animal Model; Animals; Behavior; Chromosome Mapping; Defect; Disease; Event; Exhibits; Genes; Genetic Models; Genetic Screening; Growth; Impairment; Ion Channel; Lead; Locomotion; Missense Mutation; Molecular; Molecular Analysis; Mutation; Natural regeneration; Nerve; Neurons; Pattern; Peripheral; Play; Process; Regenerative capacity; Role; SCN8A gene; Sodium Channel; Temperature; Work; Zebrafish; cholinergic neuron; exome sequencing; genetic analysis; innovation; mutant; nerve supply; novel; regenerative; tissue regeneration; tissue repair; voltage

PROJECT SUMMARY/ABSTRACT Regenerative capacity is widespread throughout almost all animal phyla, but the distributing pattern appears to be inexplicable. This diverse regenerative distribution raises questions of how animals evolve toward loss or gain of regenerative capacity and of what cellular and molecular mechanisms control regenerative ability. One feature of regeneration is that innervation is essential for peripheral tissue regeneration. Previous studies have shown that nerves are involved in multiple regeneration processes from early to late regenerative events and that distinct neuronal subtypes, such as cholinergic and sensory neurons, play different roles during tissue regeneration. However, whether neuronal excitation is required for tissue regeneration and which neuronal subtypes are associated with these processes remains poorly known. Obtaining a genetically amenable animal model will uniquely permit the identification of the essential neuronal subtypes and establishing their roles in tissue regeneration. Through forward genetic screening to discover novel regeneration-associated genes, we recently discovered a new zebrafish mutant exhibiting locomotion disorder and impaired fin regeneration in a temperature-dependent manner. Whole-exome sequencing and further fine genetic mapping analysis identified a missense mutation in the scn8a gene, which encodes the major neuronal voltage-gated sodium channel Nav1.6. We will develop a new paradigm for ion channel-regulated tissue regeneration. We will take advantage of the temperature sensitivity of scn8a mutation to define how scn8a mutation influences locomotion behavior and multiple regenerative processes. We will elucidate principles for neurons as essential drivers of tissue regeneration. We will address the challenge of whether neuronal Scn8a is required for locomotion and tissue regeneration and which Scn8a expressing neuronal subtypes are associated with fin regenerative processes. In addition to scn8a mutant, we will investigate the other two mutants, each of which exhibits either fin re-growth defects or impaired re-patterning, to uncover unidentified regeneration-associated genes and underlying mechanisms. The proposed study will construct genetic models for tissue regeneration, leading to the discovery of valuable genes regulating tissue regeneration and establishment of regenerative networks.

项目总结/摘要 再生能力在几乎所有的动物门中都是普遍存在的，但其分布模式 似乎无法解释这种多样的再生分布提出了动物如何进化到 再生能力的丧失或获得，以及控制再生能力的细胞和分子机制。 再生的一个特征是神经支配对于外周组织再生是必不可少的。以前的研究 已经表明神经参与从早期到晚期再生事件的多个再生过程 不同的神经元亚型，如胆碱能神经元和感觉神经元，在组织发育过程中发挥不同的作用， 再生然而，组织再生是否需要神经元的兴奋以及哪种神经元的兴奋是必需的， 与这些过程相关的亚型仍然知之甚少。获得一种基因上顺从的动物 该模型将唯一允许识别基本神经元亚型，并确定它们在神经元中的作用。 组织再生通过正向遗传筛选发现新的再生相关基因， 最近发现了一种新的斑马鱼突变体，表现出运动障碍和鳍再生受损， 温度依赖的方式。全外显子组测序和进一步的精细遗传图谱分析确定 编码主要神经元电压门控钠通道的scn 8a基因的错义突变 导航1.6。我们将开发一种新的离子通道调控组织再生的范例。我们将利用 scn 8a突变的温度敏感性，以确定scn 8a突变如何影响运动行为 和多种再生过程。我们将阐明神经元作为组织的基本驱动力的原则 再生我们将讨论神经元Scn 8a是否是运动和组织所必需的挑战。 Scn 8a表达神经元亚型与鳍再生过程相关。在 除了scn 8a突变体，我们将研究另外两个突变体，每个突变体都表现出鳍的再生长， 缺陷或受损的重新模式，以揭示未识别的再生相关基因和潜在的 机制等这项拟议中的研究将构建组织再生的遗传模型， 调节组织再生和建立再生网络的有价值的基因。