Discovering Mechanisms Underlying the Formation of a Straight Body Axis

发现直体轴形成的机制

• 批准号: 10390186
• 负责人: Zoe Irons
• 金额: $ 4.49万
• 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/10390186
• 关键词: Address; Anatomy; Animal Model; CRISPR/Cas technology; Cells; Cellular biology; Central cord canal structure; Cerebrospinal Fluid; Cilia; Collagen; Data; Defect; Deposition; Development; Developmental Biology; Disease; Dorsal; Drug usage; Embryo; Ensure; Event; Failure; Fertilization; Financial compensation; Fluorescent Antibody Technique; Generations; Genes; Genetic; Genetic Transcription; Genome; Growth; Growth and Development function; Head; Hour; Idiopathic scoliosis; Image; Larva; Maintenance; Masks; Modeling; Molecular; Morphogenesis; Morphology; Movement; Muscle; Muscle Contraction; Mutagenesis; Neurons; Neuropeptides; Nonsense-Mediated Decay; Organism; Pathway interactions; Pattern; Peptides; Pharmacotherapy; Phenotype; Play; 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; base; biological research; body position; cerebrospinal fluid flow; cilium motility; developmental disease; experimental study; gain of function; genetic manipulation; innovation; insight; mRNA Transcript Degradation; mutant; notochord; novel; prevent; scoliosis; 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.

项目总结/摘要： 在发育过程中，线性的头-尾体轴是如何形成的，然后在生长过程中保持不变， 明白轴向矫直缺陷-形成线性轴的形态发生过程- 导致结构性出生缺陷和脊柱弯曲，如脊柱侧凸。在这个提议中，我们使用了易处理的 斑马鱼系统，以发现基本原则的形成和维护的线性体轴。斑马鱼 胚胎最初在卵黄球周围向腹侧弯曲，但在受精后约32小时， 轴已经变直，产生了一个自主游泳的幼虫。轴向拉直需要运动的纤毛， 其在中央管中搏动并产生脑脊液（CSF）流。纤毛跳动结果 在一个失败的拉直，与幼虫保持腹曲。使用新的双突变系，在Aim 1中， 我们确定了神经肽Urp 1和Urp 1下游纤毛运动功能的机制。 这将严格测试轴向伸直依赖于尾加压素肽的模型，并建立 脊柱侧凸的新动物模型。接下来，我们评估如何形态发生运动，拉直轴停止 精确地说是在产生直轴的时候。在PKD2突变胚胎中，这种停止过程失败，导致 异常的背曲在目标2和3中，我将确定Pkd2在以下方面发挥作用的机制： 控制轴向线性。在目标2中，我将检验Pkd2调节纤毛独立过程的假设， 控制背部肌肉的收缩，为形态发生的身体运动提供力量。在目标3中，我 将测试脊索的作用，一个僵硬的杆下的中心机构，在保持直线，一旦它是 产生的，和Pkd2在这个组织中的潜在作用。总的来说，我的工作将发现新的原则， 多个组织-中央管内的运动纤毛/CSF/神经元界面，背体肌，和 脊索坐标来生成线性体轴。这项工作将对理解 一个有机体的正确解剖结构是如何通过自组织过程产生的， 了解结构性出生缺陷的起源和再生医学。我的工作将使用一个范围 创新技术，并为我提供了细胞和发育生物学方面的严格培训。