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: 10650801
• 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/10650801
• 关键词: 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.

项目摘要/摘要 再生能力普遍存在于几乎所有的动物门，但其分布格局 似乎令人费解。这种多样化的再生分布引发了动物如何进化到 再生能力的丧失或增加以及控制再生能力的细胞和分子机制。 再生的一个特点是神经支配对于周围组织再生是必不可少的。以前的研究 已经证明神经参与了从早期到晚期的多个再生过程 不同的神经元亚型，如胆碱能神经元和感觉神经元，在组织中扮演不同的角色 再生。然而，组织再生是否需要神经元兴奋，以及哪个神经元 与这些过程相关的亚型仍然鲜为人知。获得一种遗传上可服从的动物 模型将唯一地允许识别基本的神经元亚型并确定它们在 组织再生。通过正向遗传筛选发现新的再生相关基因，我们 最近发现一种新的斑马鱼突变体，表现出运动障碍和鳍再生障碍 依赖于温度的方式。全外显子组测序和进一步的精细遗传作图分析 编码主要神经元电压门控钠通道的scn8a基因的错义突变 Nav1.6.我们将为离子通道调节的组织再生开发一种新的范例。我们会利用这个优势 研究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基本驱动力的原理 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 再生和表达神经元亚型的Scn8a与FIN再生相关。在……里面 除了scn8a突变体外，我们还将研究另外两个突变体，每个突变体都表现出鳍的重新生长。 缺陷或受损的重新构图，以揭示未知的再生相关基因和潜在的 机制。这项拟议的研究将构建组织再生的遗传模型，从而导致这一发现 有价值的基因调控组织再生和再生网络的建立。