Maintenance, germ-layer induction, and patterning of neuromesodermal progenitors

神经中胚层祖细胞的维持、胚层诱导和模式化

• 批准号: 10416020
• 负责人: Benjamin L Martin
• 金额: $ 32.95万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-05 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10416020
• 关键词: Affect; Anterior; Binding; Biological Assay; Blood Vessels; Bone Morphogenetic Proteins; Brain; Cell Culture Techniques; Cell Fate Control; Cell Lineage; Cell Maintenance; Cell Transplantation; Cells; ChIP-seq; Competitive Binding; Complement; Data; Development; Dorsal; Ectoderm; Embryo; Embryonic Development; Epidermis; Fluorescence; Genes; Germ Layers; Goals; Growth; Head; Heat-Shock Response; Human; Individual; Knowledge; Laboratories; Light; Maintenance; Mesoderm; Mesoderm Cell; Methods; Microscopy; Molecular; Mus; Muscle; Muscle Cells; Neurons; Outcome; Output; Paper; Pathway interactions; Pattern; Population; Positioning Attribute; Process; Property; Proteins; Publishing; Regenerative Medicine; Regulation; Resolution; Rest; Role; Signal Pathway; Signal Transduction; Signaling Protein; Skeletal Muscle; Somites; Source; Specific qualifier value; Spinal Cord; System; Testing; Time; Tissues; Transgenic Organisms; Undifferentiated; WNT Signaling Pathway; Work; Zebrafish; base; beta catenin; blastomere structure; bone; cell motility; cell type; combinatorial; gastrulation; genetic manipulation; in vivo; insight; morphogens; mutant; novel; progenitor; relating to nervous system; stem cell biology; stem cells; transcription factor; transcriptome sequencing; transgene expression; vertebrate embryos

During the establishment of the vertebrate body, the head forms first and the rest of the body grows progressively away from the head. A population of neuromesodermal progenitor cells (NMPs) located at the posterior-most end of the embryo fuels this process of posterior growth. NMPs maintain germ-layer plasticity after the end of gastrulation, and contribute to the growing spinal cord, somites, and blood vessels. NMPs are critical cells required for the formation of the body, yet due to their late temporal role in embryogenesis and the fact that they utilize many of the same genes and signals that are essential for gastrulation, they have been extremely difficult to study. My laboratory has developed methods using the zebrafish embryo to manipulate NMPs in vivo, using a combination of cell transplantation and temporal genetic manipulations. These methods allowed us to determine the existence of zebrafish NMPs and to define some of the basic molecular properties that facilitate their germ-layer decision between mesoderm and ectoderm. This proposal builds on our past studies and will examine the molecular control of NMP maintenance and patterning. In Aim 1 we will use cell transplantation of new sox2 and sox3 mutant zebrafish lines and heat-shock inducible sox2 and Wnt signaling transgenic lines to determine how Sox2 and Wnt signaling collaborate to maintain NMPs through regulation of a new class of Wnt/βcat target genes. In Aim 2, we will determine how the Bone Morphogenetic Protein (BMP) pathway acts as a morphogen to pattern the mesodermal germ layer into distinct cell types. This aim will use new methods to precisely regulate the intensity and duration of signaling pathway activation in individual transplanted cells. The results of our study will help decipher some of the basic underpinnings of vertebrate body formation, and will provide essential information regarding the use of stem cells for regenerative medicine.

在脊椎动物身体的建立过程中，头部首先形成，身体的其余部分 逐渐远离头部生长神经中胚层祖细胞（NMPs） 位于胚胎的最后端，为后面的生长过程提供了燃料。NMPs 在原肠胚形成结束后保持胚层可塑性，并有助于生长的脊髓 脊髓体节和血管NMP是身体形成所需的关键细胞， 然而，由于它们在胚胎发生中的晚期作用，以及它们利用许多 同样的基因和信号是原肠胚形成所必需的，他们已经非常困难， study.我的实验室已经开发出使用斑马鱼胚胎在体内操纵NMPs的方法， 使用细胞移植和时间遗传操作的组合。这些方法 使我们能够确定斑马鱼核基质蛋白的存在，并定义一些基本的分子， 促进它们在中胚层和外胚层之间的胚层决定的特性。这项建议 建立在我们过去的研究，并将检查NMP维持的分子控制， 模式化在目标1中，我们将使用新的sox 2和sox 3突变斑马鱼系的细胞移植 以及热休克诱导的sox 2和Wnt信号转导转基因系，以确定Sox 2和Wnt如何 信号传导通过调节一类新的Wnt/βcat靶基因来协同维持NMP。 在目标2中，我们将确定骨形态发生蛋白（BMP）通路如何作为形态发生素， 使中胚层形成不同的细胞类型。这一目标将使用新的方法， 精确调节个体中信号通路激活的强度和持续时间， 移植细胞我们的研究结果将有助于破译一些基本的基础， 脊椎动物身体的形成，并将提供有关干细胞的使用基本信息 用于再生医学