Genetic and Cellular Analysis of Vertebrate Anterior Neurulation

脊椎动物前神经的遗传和细胞分析

基本信息

批准号：
9351708
负责人：
JENNIFER O LIANG
金额：
$ 45.48万
依托单位：
UNIVERSITY OF MINNESOTA DULUTH
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9351708
关键词：
Adherens Junction Anencephaly Anterior Apical Authorship Basic Science Biochemical Biological Assay Biology Birth Brain CRISPR/Cas technology Cadherins Candidate Disease Gene Cell Adhesion Cell Polarity Cell division Cells Cellular Morphology Cessation of life Complex Congenital Abnormality Data Defect Development Diagnosis Embryo Environment Environmental Risk Factor Epithelium Failure Fibroblast Growth Factor Gene Proteins Gene Targeting Generations Genes Genetic Goals Head Human Injectable Laboratories Lateral Link Mammals Manuscripts Measures Medial Mesenchyme Mesoderm Modeling Mutation Neural Tube Closure Neural Tube Defects Neural tube Neuroectoderm Nodal Nutritional Pathway interactions Phenotype Process Proteins Publications Research Role Signal Pathway Signal Transduction Spinal Cord Students Techniques Technology Testing Tight Junctions Time Tissues WNT Signaling Pathway Work Zebrafish base brain abnormalities cranium differential expression experience experimental study falls genome editing graduate student insight laboratory curriculum loss of function mutant nectin neural plate neuroepithelium prevent transcriptome transcriptome sequencing undergraduate student

项目摘要

PROJECT SUMMARY Neural tube defects (NTD), resulting from a failure in the transformation of a flat neural plate into a closed neural tube, occur in approximately one in every thousand births. When the anterior neural tube fails to close, it results in a fatal birth defect called anencephaly in which the brain and skull fail to develop. Our long- term goal is to use zebrafish to define the genetic and cellular mechanisms that control closure of the anterior neural tube. We found that zebrafish embryos with reduced Nodal signaling have an open anterior neural tube phenotype analogous to anencephaly. Our previous research demonstrated that the role of Nodal signaling is to induce anterior mesodermal and mesendodermal tissues. These tissues then communicate with the overlying neuroepithelium in some way that promotes neural tube closure. The experiments in this proposal will define the steps downstream of Nodal signaling and mesoderm/mesendoderm formation that are required for anterior neurulation. A recent RNAseq screen in the Liang laboratory identified over 3,000 genes that are differentially expressed between embryos with open and closed neural tubes. Many of these genes fall into pathways are good candidates to be working with Nodal signaling to promote neural tube closure. The role of these candidate pathways will be tested through loss-of- function and rescue experiments that will determine whether they are necessary for neural tube closure and whether they interact with Nodal signaling. The second part of this proposal will define defects in the neuroepithelium that prevent it from closing. Biochemical and cellular techniques that measure cell polarity, cell morphology, and protein localization and expression will define the changes that correlate with the open neural tube phenotype. These research goals will be tightly linked to the creation of an “Experimental Biology” course that will increase the opportunities for undergraduate students to participate in hypothesis-driven, original research. In this course, small groups of undergraduate students will work together to generate a hypothesis on the role of one of the RNAseq candidate genes in anterior neurulation. They will then test their hypothesis by characterizing the neural tube phenotype of a new zebrafish mutant they make using CRISPR/Cas9 genome editing. Anterior neurulation in mammals requires a similar tissue, the head mesenchyme, that also underlies the developing neural tube. Thus, this cooperative research by undergraduate and graduate students has great potential to reveal new genes and signaling pathways that could underlie anencephaly in humans. !

项目摘要神经管缺陷（NTD），是由于扁平神经板转换为一个封闭的神经管，每千生中大约有一个。当前神经管无法结束时，它导致致命的先天缺陷称为事件，其中大脑和头骨无法发育。我们的长期术语目标是使用斑马鱼来定义控制前部关闭的遗传和细胞机制神经管。我们发现斑马鱼胚胎具有降低的淋巴结信号传导的前神经管表型类似于本次域。我们以前的研究表明，淋巴结信号的作用是诱导前中胚层和中胚层组织。然后这些组织与以某种方式上覆盖神经冲突，可促进神经管道闭合。该提案中的实验将定义节点信号传导下游的步骤和前神经化所需的中胚层/中胚层形成。最近的RNASEQ屏幕梁实验室确定了3,000多个基因，这些基因在开放和开放和闭合神经管。这些基因中的许多属于途径是与Nodal一起工作的好候选者信号传导以促进神经元管闭合。这些候选途径的作用将通过损失进行测试功能和救援实验将确定它们是否需要神经管闭合和它们是否与淋巴结信号相互作用。该提案的第二部分将在神经皮质可防止其关闭。测量细胞极性，细胞的生化和细胞技术形态学，蛋白质定位和表达将定义与开放中性相关的变化管表型。这些研究目标将与创建“实验生物学”课程的创建紧密相关，该课程将增加本科生参加假设驱动的原始研究的机会。在本课程，一小群本科生将共同努力，以产生关于前神经化中的RNASEQ候选基因之一。然后，他们将通过表征他们使用CRISPR/CAS9基因组制造的新斑马鱼突变体的神经管表型编辑。哺乳动物的前神经化需要类似的组织，即头部杂物，这也是基础的发育中的神经管。那是本科生和研究生的合作研究很棒潜力揭示可能在人类中本脑的基础的新基因和信号传导途径。呢