Neuroskeletal Systems Biology in Zebrafish

 描述(由申请人提供)：使用简单的脊椎动物模型来确定神经肌肉调节骨骼的分子和细胞基础，代表了一种强大但尚未探索的策略，以揭示高等生物中的同源路径。在此背景下，体内筛选和快速基因敲除策略具有独特的潜力来识别骨外调控成骨的途径，然而在传统的体内骨合成代谢模型中，这些策略基本上是无法实现的。为了克服这一障碍，在这个项目中，我们将开发再生斑马鱼尾鳍，这是一种膜内成骨模型，概括了哺乳动物骨形成的主要阶段，作为发现神经骨骼途径的快速遗传平台。特别是，我们的中心目标是将再生鳍中的定量骨成像与斑马鱼击倒和筛选策略相结合，以识别骨生长、图案化和矿化的新神经调节因子。如果成功，这些研究将建立强大的体内分析方法，用于评估再生鳍中的神经骨骼途径，开发用于测量这一过程中骨骼生长和矿化的技术工具箱，并确定神经骨骼发现的有价值的化学切入点。通过这样做，该项目将推动再生斑马鱼的鳍成为体内的一种新的 为新兴的神经骨骼系统生物学领域建立模型，并在斑马鱼骨骼中催化更广泛的途径和治疗性筛选工作。这项研究将在高度协作的咨询委员会(其集体成果包括再生生物学、斑马鱼骨骼发育、神经骨骼生物学和进化基因组学的开创性发现)的指导下进行，提供丰富的多学科研究经验。在他过渡到一个新的研究领域时，通过提供PI技术培训、专家指导和职业指导，该项目将直接促进他向研究独立的发展。

 DESCRIPTION (provided by applicant): The use of simple vertebrate models to identify the molecular and cellular basis of neuromuscular regulation of bone represents a powerful yet unexplored strategy to uncover homologous pathways in higher organisms. In this context, in vivo screens and rapid gene knockdown strategies hold unique potential to identify extraskeletal pathways regulating osteogenesis, however such strategies are largely inaccessible in traditional in vivo models of bone anabolism. Toward overcoming this hurdle, in this project we will develop the regenerating zebrafish tail fin, a model of intramembranous ossification that recapitulates the major phases of mammalian bone formation, as a rapid genetic platform for neuroskeletal pathway discovery. In particular, our central objective is to integrate quantitative bone imaging in the regenerating fin with zebrafish knockdown and screening strategies to identify novel neural regulators of bone outgrowth, patterning, and mineralization. If successful, these studies will establish powerful in vivo assays for neuroskeletal pathway assessment in the regenerating fin, develop the technological toolbox for measuring bone growth and mineralization in this process, and identify valuable chemical entry points for neuroskeletal discovery. In doing so, this project will advance the regenerating zebrafish fin as a novel in vivo model for the emerging field of neuroskeletal systems biology, and catalyze broader pathway and therapeutic screening efforts in the zebrafish skeleton. This research will be carried out under the mentorship of a highly collaborative advisory committee (whose collective achievements include seminal findings in regenerative biology, zebrafish skeletal development, neuroskeletal biology, and evolutionary genomics), providing a rich, multidisciplinary research experience. By providing the PI technical training, expert guidance, and career mentorship as he transitions to a new research field, this project will directly facilitate his progression to research independence.