Discovering Mechanisms Underlying the Formation of a Straight Body Axis

发现直体轴形成的机制

• 批准号: 10621160
• 负责人: Zoe Irons
• 金额: $ 4.62万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-16 至 2024-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10621160
• 关键词: Address; Anatomy; Animal Model; CRISPR/Cas technology; Cells; Cellular biology; Central cord canal structure; Cerebrospinal Fluid; Cilia; Collagen; Compensation; Data; Defect; Deposition; Development; Developmental Biology; Disease; Dorsal; Drug usage; Embryo; Ensure; Event; Failure; Fertilization; Generations; Genes; Genetic; Genetic Transcription; Genome; Growth; Growth and Development function; Head; Hour; Idiopathic scoliosis; Image; Immunofluorescence Immunologic; Larva; Maintenance; Modeling; Molecular; Morphogenesis; Morphology; Movement; Muscle; Muscle Contraction; Mutagenesis; Neurons; Neuropeptides; Nonsense-Mediated Decay; Organism; Pathway interactions; Pattern; Peptides; Pharmacotherapy; Phenotype; Process; RNA; Regenerative Medicine; Research; Research Personnel; Rod; Role; SHH gene; Scanning Electron Microscopy; Science; Shapes; Signal Transduction; Source; Spinal; Stereotyping; Structural Congenital Anomalies; Structure; Swimming; System; Tail; Techniques; Testing; Tissues; Training; Vertebral column; Vertebrates; Work; Zebrafish; biological research; body position; cerebrospinal fluid flow; cilium motility; developmental disease; experimental study; gain of function; genetic manipulation; innovation; insight; loss of function; mRNA Transcript Degradation; model organism; mutant; notochord; novel; prevent; scoliosis; self organization; smoothened signaling pathway

PROJECT SUMMARY/ ABSTRACT: How the linear head-to-tail body axis forms during development and is then maintained during growth is little understood. Defects in axial straightening — the morphogenetic process through which a linear axis forms — result in structural birth defects and spinal curves such as scoliosis. In this proposal, we use the tractable zebrafish system to discover principles underlying the formation and maintenance of a linear body axis. Zebrafish embryos are initially curved ventrally around a ball of yolk but, by around 32 hours post fertilization, the embryonic axis has straightened to produce an autonomously swimming larva. Axial straightening requires motile cilia, which beat and generate cerebrospinal fluid (CSF) flow in the central canal. Abnormalities in cilia beating result in a failure of straightening, with larvae maintaining ventral curves. Using a novel double mutant line, in Aim 1 we determine mechanisms underlying the function of neuropeptides Urp1 and Urp1 downstream of cilia motility. This will rigorously test the model that axial straightening depends on Urotensin peptides as well as establish new animal models for scoliosis. Next, we assess how morphogenetic movements that straighten the axis stop precisely when a straight axis is generated. In pkd2 mutant embryos, this stopping process fails, leading to abnormal dorsal curves. In Aims 2 and 3, I will determine the mechanisms through which Pkd2 functions in controlling axial linearity. In Aim 2, I will test the hypothesis that Pkd2 regulates cilia-independent processes that control the dorsal muscle contractions which supply the force for morphogenetic body movements. In Aim 3, I will test the role of the notochord, a stiff rod down the center of the body, in maintaining straightness once it is generated, and the potential role of Pkd2 in this tissue. Overall, my work will discover new principles of how multiple tissues — the motile cilia/CSF/neuronal interface within the central canal, dorsal somitic muscle, and the notochord — coordinate to generate a linear body axis. This work will have implications for understanding how the correct anatomy of an organism is generated through self-organizing processes, something important for understanding the origin of structural birth defects and for regenerative medicine. My work will use a range of innovative techniques and provide me with a rigorous training in cell and developmental biology.

项目摘要/摘要： 在开发过程中，线性从头到尾的身体轴是如何形成的，然后在生长过程中保持几乎没有 理解。轴向拉直的缺陷 - 形态学过程，线性轴形成 - 导致结构性先天缺陷和脊柱曲线，例如脊柱侧弯。在此提案中，我们使用可处理的 斑马鱼系统发现线性轴的形成和维护的基础。斑马鱼 胚胎最初是在蛋黄的球周围腹侧弯曲的，但在受精后大约32小时，胚胎 轴已伸直以产生自​​主游泳幼虫。轴向拉直需要纤毛， 在中央运河中击败并产生脑脊液（CSF）流动。纤毛击败结果异常 在拉直的失败中，幼虫保持腹侧曲线。在AIM 1中使用新颖的双重突变线 我们确定神经肽URP1和CILIA运动下游功能功能的机制。 这将严格测试轴向拉直取决于尿素蛋白辣椒的模型以及已建立的 脊柱侧弯的新动物模型。接下来，我们评估形态发生运动如何拉直轴停止 当产生直轴时。在PKD2突变胚胎中，此停止过程失败，导致 异常背曲线。在目标2和3中，我将确定PKD2在 控制轴向线性。在AIM 2中，我将检验以下假设：PKD2调节与纤毛独立的过程有关 控制为形态发生身体运动提供力的背部肌肉收缩。在AIM 3中，我 将测试Notochord的角色，即身体中心的僵硬的杆，一旦保持直率 产生，PKD2在该组织中的潜在作用。总的来说，我的工作将发现如何 多个时机 - 中央管中的纤毛/CSF/神经元界面，背肌肌肉和 脊索 - 坐标以生成线性体轴。这项工作将对理解有影响 如何通过自组织过程产生生物体的正确解剖结构，这是重要的 了解结构性先天缺陷和再生医学的起源。我的工作将使用范围 创新技术，并为我提供了细胞和发育生物学的严格培训。