Cell cycle dependent mechanisms triggering lumen formation in vivo

触发体内管腔形成的细胞周期依赖性机制

• 批准号: 10322191
• 负责人: Heidi Hehnly
• 金额: $ 30万
• 依托单位: SYRACUSE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322191
• 关键词: Ablation; Address; Animal Model; Apical; Attention; Biology; Cardiac; Cell Cycle; Cell Differentiation process; Cell Nucleus; Cell division; Cell membrane; Cells; Cellular biology; Centrosome; Cilia; Congenital Heart Defects; Cyst; Cystic Fibrosis Transmembrane Conductance Regulator; Cytokinesis; Cytosol; Defect; Development; Developmental Biology; Embryonic Development; Epithelial; Epithelial Cells; Event; Excision; Genes; Hair; Heart; Human; In Vitro; Kidney; Lasers; Left; Link; Liver; Membrane; Membrane Protein Traffic; Mesenchymal; Mesenchymal Stem Cells; Microtubules; Mitosis; Modeling; Molecular; Morphogenesis; Organ; Organelles; PLK1 gene; Pancreas; Pathway interactions; Patients; Pattern; Play; Positioning Attribute; Process; Publications; Resolution; Role; Signal Transduction; Site; System; Testing; Time; Tissues; Vertebrates; Vesicle; Work; Zebrafish; apical membrane; base; ciliopathy; cilium biogenesis; cilium motility; daughter cell; epithelial to mesenchymal transition; fluid flow; in vivo; kinetosome; optogenetics; precursor cell; premature; protein expression; trafficking; vesicle transport

SUMMARY STATEMENT: In humans and other vertebrates, motile cilia located in an organ of asymmetry play an important role in cardiac left-right development. Evidence from model organisms, such as in zebrafish organ of asymmetry, (Kupffer’s Vesicle, KV) indicates that conserved cilia-driven leftward flow establishes left- right signals to regulate target genes to control asymmetric heart morphogenesis. While events downstream of leftward flow have received much attention, little is known about how the organ of asymmetry is formed and the biology of the ciliated cells that generate fluid flow. This project addresses the broad question: How do ciliated cells develop into a functional polarized organ? To address this question we are bringing together cell biology and developmental biology to investigate how the cytokinetic bridge establishes apical polarity and a lumen in vivo. We propose that this occurs through a sequential process that starts with cell division and placement of the cytokinetic midbody, which marks a site for where the apical membrane should be placed. Cytokinetic bridge resolution (a.k.a. abscission) results in the separation of two daughter cells following mitosis allowing for the cell to initiate ciliogenesis. This process has important implications in embryogenesis, and broad implications in the role of cytokinesis in developing cellular diversity. While abscission has been examined in vitro, little has been done to examine the role of cytokinesis/abscission in epithelial establishment and de novo lumen formation in vivo. Thus, our work will test the overall hypothesis that cell division is an essential process that initiates lumen formation, ciliogenesis, and subsequently tissue morphogenesis. Here we propose to examine in the zebrafish KV a requirement for abscission in the transition of progenitor-mesenchymal-like migratory cells to epithelial-ciliated cells (tested in Aim 1). For instance, does cell division trigger KV-specific apical polarity protein expression and does division contribute to how cells are patterned to form a KV? We propose that following cytokinesis, daughter cells stay interconnected by a cytokinetic bridge while apical polarity is established. This process requires targeted membrane traffic into the cytokinetic bridge. During this time, the two daughter cells position themselves so that the cytokinetic bridge is placed where an apical membrane and lumen will form. Once the bridge is cleaved, a lumen is initiated (Aim 2) and KV cells can form primary cilia (Aim 3). We will use photoconversion to track cell fate following division, and laser ablation or optogenetics to determine whether abscission timing is important for apical polarity, cilia formation, and lumenogenesis. These studies will identify important mechanisms for de novo tissue morphogenesis.

摘要声明：在人类和其他脊椎动物中，苏里亚的母亲位于不对称器官中 在心脏左右发展中发挥重要作用。来自模型生物的证据，例如斑马鱼 不对称器官（Kupffer的囊泡，KV）表明，保守的纤毛驱动的向左流有左 - 右信号调节靶基因以控制不对称的心形形态发生。而事件下游 左流已经引起了很多关注，对如何形成不对称器官几乎了解 产生液体流动的纤毛细胞的生物学。该项目解决了一个广泛的问题：纤毛如何 细胞发展成功能性极化器官？为了解决这个问题，我们将细胞生物学汇总在一起 和开发生物学，以研究细胞力学桥如何建立顶极性和腔内 体内。我们提出，这是通过一个从细胞分裂和位置开始的顺序过程发生的 细胞动力学中体，标志着应放置顶膜的位置。细胞动力学 桥分辨率（又称脱落）导致有丝分裂后两个子细胞分离 启动纤毛生成的细胞。这个过程对胚胎发生具有重要意义，并且广泛 细胞因子在发展细胞多样性中的作用的意义。虽然已经检查了脱落 体外，几乎没有做过细胞因子/脱落在上皮机构和从头开始的作用 体内管腔形成。这是我们的工作将测试整体假设，即细胞分裂是必不可少的 引发管腔形成，纤毛发生和随后的组织形态发生的过程。我们在这里提出 在斑马鱼KV中检查祖子 - 间充质样过渡中脱落的要求 迁移到上皮细胞的细胞（在AIM 1中测试）。例如，细胞分裂会触发KV特异性 顶极性蛋白表达，分裂是否有助于细胞形成形成形成KV？我们 提出的提议是在细胞因子之后，子细胞保持互联桥的互连，而顶端 建立了极性。此过程需要靶向膜流入细胞动力学桥梁。在此期间 时间，两个子细胞将自己定位为 膜和管腔将形成。一旦桥梁裂开，就开始腔（AIM 2），KV细胞可以形成 初级纤毛（目标3）。我们将使用光转换来跟踪分裂后细胞脂肪，并激光消融或 光遗传学以确定脱落时间是否对顶极性，纤毛形成和 流明发生。这些研究将确定从头组织形态发生的重要机制。