Morphogenetic mechanisms regulating directed cell migration required to form the vertebrate posterior body

调节形成脊椎动物后体所需的定向细胞迁移的形态发生机制

• 批准号: 9982378
• 负责人: David Kimelman
• 金额: $ 33.23万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9982378
• 关键词: Adopted; Anterior; Biological Assay; Biological Models; Cell Proliferation; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Congenital Abnormality; Development; Down-Regulation; Embryo; Embryo Deaths; Failure; Gene Expression; Goals; Growth; Homeobox Genes; Individual; Lead; Measures; Mesoderm; Mesoderm Cell; Movement; Muscle; Neural tube; Neurons; Population; Process; Regulator Genes; Resolution; Somites; Spinal Cord; System; Testing; Time; Transgenic Organisms; Zebrafish; base; cell motility; cell type; cellular imaging; directional cell; epithelial to mesenchymal transition; high resolution imaging; in vivo; in vivo imaging; innovation; insight; loss of function; migration; mutant; novel; prevent; progenitor; relating to nervous system; rho; somitogenesis; stem cells

PROJECT SUMMARY Much of the vertebrate embryonic body forms from a recently discovered neuromesodermal progenitor cell population located at the most posterior end of the early embryo, in a region called the Progenitor Zone (PZ). The PZ gradually releases mesodermal cells that populate the somites (primarily muscle) as well as the neural cells that form the spinal cord, until the complete anterior-posterior axis of the embryo has been established. While the mechanisms controlling the differentiation of the neuromesodermal cells are increasingly understood, how these cells regulate the completion of the epithelial to mesenchymal transition (EMT) and directional migration from the PZ into the body largely remains a mystery, yet this process is essential for the embryo to form its anterior-posterior axis correctly. Based on our analysis of a zebrafish mutant that specifically disrupts the migration of mesodermal cells from the PZ, we have identified a unique set of PZ-expressed Progenitor Cytoskeletal Regulatory Genes (PCRGs) that we propose must be down-regulated for cells to complete the EMT and migrate from the PZ into the presomitic mesoderm (PSM). Using a novel explant assay we recently developed that allows in vivo imaging of the migrating cells at very high resolution, in Aim 1 we will determine how the PCRGs are used to regulate the directional movement of cells into the PSM using a combination of gain and loss of function studies, as well as examining how the PCRGs control Rho activity. In Aim 2 we will determine how the posterior hox genes, which through unknown mechanisms control the orderly movement of cells from the PZ into the embryonic body, specifically regulate cell movements using our novel explant system to examine directional cell migration and protrusive activity of the hox-expressing cells. We will test the hypothesis that the hox genes act to sustain the expression of the PCRGs, and thereby regulate the timing of cell entry into the PSM. Collectively, these studies will examine the premise that the migration of cells from the PZ into the PSM is regulated by the action of the PCRGs and posterior hox genes, which acting together control the orderly anteriorward migration of newly differentiating mesodermal cells, thus allowing the vertebrate embryonic body to form with remarkable fidelity. With the ease of making transgenic lines that allow temporally controlled expression of the PCRGs and hox genes as well as CRISPR mutant lines, combined with our innovative explant system for high resolution imaging, zebrafish is an excellent system for understanding the mechanisms that control the early formation of the vertebrate body.

项目摘要 许多脊椎动物的胚胎体是由最近发现的神经中胚层祖细胞形成的 位于早期胚胎最后端的群体，在一个称为祖细胞区（PZ）的区域。 PZ逐渐释放中胚层细胞，这些细胞填充体节（主要是肌肉）以及神经节。 细胞形成脊髓，直到胚胎的完整前后轴已经建立。 虽然控制神经中胚层细胞分化的机制越来越多 了解这些细胞如何调节上皮向间充质转化（EMT）的完成， 从PZ到身体的定向迁移在很大程度上仍然是一个谜，但这个过程对于 胚胎形成其前后轴正确。 基于我们对斑马鱼突变体的分析，该突变体特异性地破坏了中胚层细胞从 在PZ中，我们已经鉴定了一组独特的PZ表达的祖细胞骨架调节基因（PCRG）， 我们认为，细胞完成EMT并从PZ迁移到PZ必须下调。 有丝分裂前中胚层（PSM）。使用我们最近开发的一种新的外植体测定法， 在目标1中，我们将确定如何使用PCRG来调节细胞的迁移。 使用功能获得和丧失研究的组合将细胞定向移动到PSM中，以及 研究PCRG如何控制Rho活性。 在目标2中，我们将确定后hox基因是如何通过未知的机制控制细胞增殖的。 细胞从PZ到胚胎体的有序运动，特别是使用 我们的新的外植体系统，以检查定向细胞迁移和hox表达细胞的增殖活性， 细胞我们将检验hox基因维持PCRG表达的假设， 调节细胞进入PSM的时间。 总的来说，这些研究将检查细胞从PZ迁移到PSM的前提， 由PCRG和后部hox基因的作用调节，它们共同作用控制有序的 新分化的中胚层细胞向前迁移，从而使脊椎动物胚胎 身体以非凡的保真度形成。由于容易制造允许暂时性的转基因品系， PCRG和hox基因以及CRISPR突变株系的受控表达，结合我们的 创新的外植体系统高分辨率成像，斑马鱼是一个很好的系统，了解 控制脊椎动物身体早期形成的机制。

项目成果

Anterior-posterior patterning in early development: three strategies.

• DOI: 10.1002/wdev.25
• 发表时间: 2012-03
• 期刊: Wiley interdisciplinary reviews. Developmental biology
• 作者: Kimelman D;Martin BL
• 通讯作者: Martin BL

Hedgehog and Bmp polarize hematopoietic stem cell emergence in the zebrafish dorsal aorta.

• DOI: 10.1016/j.devcel.2009.04.014
• 发表时间: 2009-06
• 期刊: DEVELOPMENTAL CELL
• 影响因子: 11.8
• 作者: Wilkinson, Robert N.;Pouget, Claire;Gering, Martin;Russell, Angela J.;Davies, Stephen G.;Kimelman, David;Patient, Roger
• 通讯作者: Patient, Roger

Bmp inhibition is necessary for post-gastrulation patterning and morphogenesis of the zebrafish tailbud.

• DOI: 10.1016/j.ydbio.2009.02.016
• 发表时间: 2009-05-01
• 期刊: DEVELOPMENTAL BIOLOGY
• 影响因子: 2.7
• 作者: Row, Richard H.;Kimelman, David
• 通讯作者: Kimelman, David

Canonical Wnt signaling dynamically controls multiple stem cell fate decisions during vertebrate body formation.

• DOI: 10.1016/j.devcel.2011.11.001
• 发表时间: 2012-01-17
• 期刊: DEVELOPMENTAL CELL
• 影响因子: 11.8
• 作者: Martin, Benjamin L.;Kimelman, David
• 通讯作者: Kimelman, David

Restricted expression of cdc25a in the tailbud is essential for formation of the zebrafish posterior body.

• DOI: 10.1101/gad.233577.113
• 发表时间: 2014-02-15
• 期刊: Genes & development
• 影响因子: 10.5
• 作者: Bouldin CM;Snelson CD;Farr GH 3rd;Kimelman D
• 通讯作者: Kimelman D