Molecular and Genetic Analysis of Fin Regeneration in Zebrafish

Parent Grant R35GM137878项目摘要 再生能力在几乎所有动物门中都很普遍，但其分布模式似乎是 令人费解。这种多样化的再生分布提出了一个问题，即动物是如何进化到失去或获得的 再生能力和什么细胞和分子机制控制再生能力。一项功能 再生的关键在于神经对周围组织的再生至关重要。先前的研究表明 神经参与了从早期到晚期的多个再生过程，而且 神经元亚型，如胆碱能神经元和感觉神经元，在组织再生过程中扮演着不同的角色。 然而，组织再生是否需要神经元兴奋，以及哪些神经元亚型是必需的 与这些过程相关的信息仍然鲜为人知。获得一种可遗传的动物模型将会 唯一允许识别基本神经元亚型并确定其在组织中的作用 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 机械装置。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。

PROJECT SUMMARY FOR PARENT GRANT R35GM137878 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.