Genetic and chemical screening in zebrafish to study tendon development

对斑马鱼进行遗传和化学筛选以研究肌腱发育

• 批准号: 8574158
• 负责人: JENNA L GALLOWAY
• 金额: $ 24.9万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-12-10 至 2015-12-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8574158
• 关键词: Address; Biological Models; Biomechanics; Cartilage; Cells; Chemicals; Complement; Development; Developmental Biology; Embryo; Embryonic Development; Gene Expression; Genes; Genetic; Genetic Screening; Goals; Histocompatibility Testing; Histologic; Injury; Libraries; Ligaments; Location; Modeling; Molecular; Morphogenesis; Motion; Movement; Muscle; Musculoskeletal System; Pathway interactions; Pattern; Population; Property; Regulation; Role; Signal Pathway; Sports; Techniques; Tendon Injuries; Tendon structure; Time; Tissues; Transcript; Transgenic Organisms; Validation; Vertebrates; Work; Zebrafish; age related; bone; chemical genetics; craniofacial; developmental genetics; improved; insight; ligament injury; loss of function; novel; repaired; screening

Tendons transmit force from muscles to bone, enabling movement, wfiile ligaments connect bone to bone, providing stabilization. Not only is the integrity and strength of the attachment important, but the location of the connections within the musculoskeletal system must be strictly regulated for efficient motion. Sports or age-related injuries to tendons or ligaments are extremely common, and although the injury can be slowly repaired, the original biomechanical properties of the tendon or ligament are never fully restored. Despite how essential these tissues are for mobility, we know very little about their molecular regulation during embryonic development. The goal of this project is to discover the pathways regulating tendon and ligament formation and differentiation in the embryo. To accomplish this, we will use the zebrafish, a powerful genetic and developmental vertebrate model system'. First, zebrafish craniofacial tendons are molecularly and histologically similar to mammalian tendons. Co-expression studies with muscle and cartilage markers demonstrate the presence of tendon transcripts at muscle-cartilage connection poi nts, thus defining a distinct population of tendon cells. To understand how zebrafish tendons form, this proposal will determine the role of distinct tissue types and known signaling pathways through loss of function analysis. Additionally, transgenic zebrafish will be generated to study tendon formation and morphogenesis over real developmental time, a technique that is unfeasible in other vertebrates. To identify novel pathways that regulate tendon development, over one thousand com pounds from a known bioactive compound library were screened and 20 putative hits were identified. Further work will focus on the validation and analysis of these hits and the pathways they target in tendon developm ent. Complementing this chemical screen, a forward genetic screen will be performed to discover and characterize new pathways regulating tendon development. These studies will directly address the major questions in the field of tendon developmental biology, and may ultimately, aid in the development of new therapies for tendon or ligament injury.

肌腱将力从肌肉传递到骨骼，使运动成为可能，而韧带将骨骼连接到骨骼， 提供稳定性。不仅是附件的完整性和强度很重要，但位置 肌肉骨骼系统内的连接必须被严格调节以实现有效的运动。体育或 与年龄有关的肌腱或韧带损伤是非常常见的，虽然损伤可以慢慢地 在修复后，肌腱或韧带的原始生物力学特性从未完全恢复。尽管 这些组织对于运动是多么重要，我们对它们在运动过程中的分子调控知之甚少。 胚胎发育这个项目的目标是发现调节肌腱和韧带的途径 胚胎的形成和分化。为了实现这一目标，我们将使用斑马鱼，一种强大的遗传基因， 和发育脊椎动物模型系统“。首先，斑马鱼颅面肌腱是分子， 在组织学上与哺乳动物的肌腱相似。肌肉和软骨标记物的共表达研究 证明了肌腱转录本在肌肉软骨连接点的存在，从而定义了一个独特的 肌腱细胞群。为了了解斑马鱼肌腱是如何形成的， 不同的组织类型和已知的信号通路通过功能丧失分析。此外，本发明还 将产生转基因斑马鱼，以研究肌腱形成和形态发生超过真实的。 这是一种在其他脊椎动物中不可行的技术。为了确定新的途径， 调节肌腱发育，从已知的生物活性化合物库中筛选出超过一千种化合物， 筛选并鉴定了20个推定的命中。进一步的工作将集中在验证和分析这些 点击和他们在肌腱发育的目标途径。补充这一化学筛选，一个前锋 将进行遗传筛选，以发现和表征调节肌腱发育的新途径。 这些研究将直接解决肌腱发育生物学领域的主要问题， 最终，有助于开发肌腱或韧带损伤的新疗法。