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：在人类和其他脊椎动物中，活动的纤毛位于不对称的器官中 在心脏左右发育中起重要作用。来自模式生物的证据，如斑马鱼 不对称器官(库普弗氏囊泡，KV)表明，保守的纤毛驱动的左向血流建立了左- 调节靶基因的正确信号，以控制不对称的心脏形态发生。而下游的事件 向左流动受到了很大的关注，关于不对称器官是如何形成的，以及 纤毛细胞产生液体流动的生物学。这个项目解决了一个广泛的问题：纤毛是如何 细胞发育成一个有功能的极化器官？为了解决这个问题，我们将细胞生物学 和发育生物学来研究细胞运动桥如何建立顶极和管腔 活着。我们认为，这是通过一个连续的过程发生的，这个过程始于细胞分裂和细胞的放置 细胞动学中体，标志着顶膜应该放置的位置。细胞动学 网桥分辨率(也称为分裂)导致有丝分裂后两个子细胞分离，从而允许 启动纤毛发生的细胞。这一过程在胚胎发育中具有重要意义，并具有广泛的应用前景 胞质分裂在发展细胞多样性中的作用。虽然对离职进行了审查，但 在体外，很少有人研究胞质分裂/脱落在上皮建立和新生中的作用。 体内的管腔形成。因此，我们的工作将检验细胞分裂是必不可少的总体假设。 启动管腔形成、纤毛形成以及随后的组织形态形成的过程。在这里，我们建议 在斑马鱼KV中研究祖细胞-间充质样细胞转变中的脱落要求 向上皮纤毛细胞迁移(在AIM 1中测试)。例如，细胞分裂是否会触发KV特异性 顶端极性蛋白的表达和分裂对细胞如何形成KV有贡献吗？我们 认为在胞质分裂后，子代细胞在顶端时通过细胞动桥保持相互连接 两极是确定的。这一过程需要靶向的膜交通进入细胞动态桥。在此期间 时间，两个子细胞自身定位，以便细胞动态桥被放置在顶端 就会形成膜和腔。一旦桥被切割，腔就会启动(目标2)，KV细胞就可以形成 初级纤毛(目标3)。我们将使用光转换来跟踪细胞分裂后的命运，以及激光消融或 光遗传学，以确定脱落时间是否对顶极、纤毛形成和 管腔形成。这些研究将确定新生组织形态发生的重要机制。