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: 10439645
• 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/10439645
• 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.

项目概要/摘要 再生能力几乎遍及所有动物门，但分布模式 似乎有些莫名其妙。这种多样化的再生分布提出了动物如何进化的问题 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。 再生的特征之一是神经支配对于周围组织再生至关重要。之前的研究 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，例如胆碱能神经元和感觉神经元，在组织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兴奋以及哪些神经元兴奋是必需的。 与这些过程相关的亚型仍然知之甚少。获得基因上顺从的动物 该模型将独特地允许识别基本的神经元亚型并确定它们在 组织再生。通过正向遗传筛选来发现新的再生相关基因，我们 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 温度依赖方式。确定了全外显子组测序和进一步的精细遗传图谱分析 scn8a 基因的错义突变，该基因编码主要神经元电压门控钠通道 导航1.6。我们将开发离子通道调节组织再生的新范例。我们将利用 scn8a 突变的温度敏感性以确定 scn8a 突变如何影响运动行为 和多个再生过程。我们将阐明神经元作为组织的重要驱动因素的原理 再生。我们将解决神经元 Scn8a 是否是运动和组织所必需的挑战 再生以及表达 Scn8a 的神经元亚型与鳍再生过程相关。在 除了 scn8a 突变体之外，我们还将研究其他两个突变体，每个突变体都表现出鳍重新生长 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。拟议的研究将构建组织再生的遗传模型，从而发现 调节组织再生和再生网络建立的有价值基因。