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

基本信息

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
项目类别：
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

项目摘要

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.

在建立脊椎动物的过程中，头部首先形成，其余的身体逐渐远离头部。神经霉菌祖细胞（NMP）的种群位于胚胎的最后端燃料会为后验生长带来。 NMP 过口结束后保持菌丝层可塑性，并有助于生长的脊柱绳索，骨和血管。 NMP是形成人体所需的关键细胞，然而，由于它们在胚胎发生中的较晚临时作用以及他们利用许多事实相同的基因和信号对于过度来说，它们非常困难学习。我的实验室已经开发了使用斑马鱼胚胎来操纵NMP的方法，结合细胞移植和暂时的遗传操纵。这些方法允许我们确定斑马鱼NMP的存在并定义一些基本分子促进中胚层和外胚层之间其细菌层决策的特性。这个建议建立在我们过去的研究的基础上，并将检查NMP维护的分子控制和图案。在AIM 1中，我们将使用新的Sox2和Sox3突变斑马鱼线的细胞移植以及热震诱导的Sox2和Wnt信号转基因线，以确定SOX2和Wnt的方式信号通过调节新类别的Wnt/βCAT靶基因来维持NMP。在AIM 2中，我们将确定骨形态发生蛋白（BMP）途径如何充当形态学将中胚层生殖层分为不同的细胞类型。这个目标将使用新方法精确调节单个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解读一些基本的基础脊椎动物的形成，并将提供有关干细胞使用的基本信息用于再生医学。