Mechanisms of microtubule-mediated cranial neural crest EMT and differentiation

微管介导的颅神经嵴EMT和分化机制

基本信息

批准号：
10633228
负责人：
Crystal D Rogers
金额：
$ 14.93万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-02 至 2025-06-01
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10633228
关键词：
Actins Adherens Junction Adhesions Adult Affect Automobile Driving Biological Assay Cadherins Cartilage Cell Adhesion Cell Adhesion Molecules Cell Polarity Cell membrane Cell-Cell Adhesion Cells Central Nervous System Cephalic Chickens Complex Congenital Abnormality Cranial Nerves Cytoskeleton Data Defect Development Disease E-Cadherin Ectoderm Ectoderm Cell Elements Embryo Enteric Nervous System Environmental Risk Factor Epidermis Epigenetic Process Epithelium Facial nerve structure Future Genetic Genetic Screening Genetic Transcription Goals Growth and Development function Health Human Human Development Human body Intermediate Filaments Invaded Link Malignant Neoplasms Mediating Membrane Methods Microfilaments Microtubules Mission Modeling Natural regeneration Neural Crest Neural Crest Cell Neural Fold Neural Inhibition Neural Tube Closure Neural tube Neuroepithelial Neuroglia Neurons Oculomotor nerve structure Pathologic Pattern Peripheral Nervous System Process Proteins Public Health Publishing Regulator Genes Research Role Signal Transduction Specific qualifier value Structure Testing Tissues Transcript Transcriptional Regulation Trigeminal System Tubulin United States National Institutes of Health Work cadherin-11 cell motility cell type craniofacial bone craniofacial development epithelial to mesenchymal transition experimental study gain of function in vivo loss of function migration nervous system disorder neural neurodevelopment neuroregulation novel marker novel strategies oculomotor physical separation premature trafficking transcription factor

项目摘要

Project Summary/Abstract Defective development of neural crest (NC) cells can cause structural and neurological disorders that have long-term deleterious effects on human health. NC cells form the neurons and glia of the peripheral and en- teric nervous systems in addition to craniofacial bone and cartilage. Early formation and separation of NC cells from the adherent neural tube is a process tightly regulated by cell signaling, epigenetic and transcrip- tional changes, and altered cell-cell adhesion via changing cadherin protein localization. Structurally, as the neural folds rise to meet in the center of the embryo to create the neural tube, non-neural ectodermal cells meet and cover the embryo, eventually differentiating into epidermis and placodes. After neural tube clo- sure, NC cells undergo an epithelial to mesenchymal transition (EMT), leave their neuroepithelial neigh- bors, and migrate throughout the embryo, invading various tissues, and creating diverse derivatives. Rapid changes at transcriptional, translational, and post-translational levels allow for dynamic transitions in cell polarity, adhesion, and migration. Several transcription factors function as regulators of genes encoding cadherin proteins during NC cell development, but there is a critical lack of information about the more rapid processes that alter cadherin protein localization during NC EMT. Specifically, we aim to understand how microtubules and related cytoskeletal factors regulate cell-cell adhesion at this developmental stage. Based on our published work and preliminary data, -III tubulin (TUBB3) is upregulated in NC cells at the onset of NC EMT. Our objective is to understand the complex mechanisms that control NC EMT and subsequent NC differentiation by defining how the microtubule element, TUBB3, regulates cadherin protein localization during NC cell EMT and differentiation in vivo. Further, we will identify how perturbation of TUBB3 affects the structure of other cytoskeletal elements during NC EMT. In Aim 1 we will perform gain and loss of TUBB3 experiments followed by quantitative analyses of changes in the expression and localization of epi- thelial and migratory cadherins (CDH2, CDH1, and CDH11), tissue-specific markers (NC and neural tube), cell migration and cranial NC differentiation using quantitative spatial approaches at the transcript and pro- tein levels in the chicken (Gallus gallus), which mirrors human development at early stages. In Aim 2, we will perform perturbations of TUBB3 to determine how defective TUBB3-mediated microtubule assembly affects the localization of microtubules, intermediate filaments, and actin filaments during NC EMT. With these experiments, we expect to integrate both traditional and novel approaches to understanding the com- plex links between cell adhesion and cytoskeletal arrangements during NC EMT. We hope to contribute a missing, fundamental element to our knowledge of the protein network that drives the development of NC cells, which is crucial for the identification of novel markers of normal and abnormal development of NC- derived adult cell types.

项目概要/摘要神经嵴 (NC) 细胞发育缺陷可导致结构性和神经系统疾病 NC 细胞形成外周神经元和神经胶质细胞，对人类健康产生长期有害影响。除颅面骨和软骨外的颅神经系统 NC 的早期形成和分离。来自贴壁神经管的细胞是一个受到细胞信号传导、表观遗传和转录严格调控的过程。结构上的改变，以及通过改变钙粘蛋白定位来改变细胞与细胞的粘附。神经褶皱上升并在胚胎中心相遇，形成神经管、非神经外胚层细胞接触并覆盖胚胎，最终分化为表皮和基板。当然，NC 细胞经历上皮间质转化 (EMT)，离开其神经上皮邻居 bors，并在整个胚胎中迁移，侵入各种组织，并产生多种快速衍生物。转录、翻译和翻译后水平的变化允许细胞内的动态转变极性、粘附和迁移等多种转录因子可作为编码基因的调节因子。 NC 细胞发育过程中钙粘蛋白的存在，但目前严重缺乏关于更快速的信息具体来说，我们的目标是了解在 NC EMT 期间改变钙粘蛋白定位的过程。微管和相关的细胞骨架因子在此发育阶段调节细胞间粘附。根据我们发表的工作和初步数据，-III 微管蛋白 (TUBB3) 在 NC 细胞中在我们的目标是了解控制 NC EMT 及其后续工作的复杂机制。通过定义微管元件 TUBB3 如何调节钙粘蛋白定位来进行 NC 分化此外，我们将确定 TUBB3 的扰动如何影响 NC 细胞 EMT 和体内分化。在目标 1 中，我们将执行 NC EMT 期间其他细胞骨架元素的结构的增益和丢失。 TUBB3实验随后对epi-表达和定位的变化进行定量分析皮细胞和迁移钙粘蛋白（CDH2、CDH1 和 CDH11）、组织特异性标记物（NC 和神经管）、使用定量空间方法在转录本和亲细胞上进行细胞迁移和颅脑 NC 分化鸡 (Gallus gallus) 中的蛋白质水平反映了人类早期发育阶段。将执行 TUBB3 的扰动以确定 TUBB3 介导的微管组装有缺陷影响 NC EMT 期间微管、中间丝和肌动蛋白丝的定位。在这些实验中，我们希望整合传统和新颖的方法来理解计算机 NC EMT 期间细胞粘附和细胞骨架排列之间的复杂联系我们希望做出贡献。我们对驱动 NC 发展的蛋白质网络的了解缺失了基本要素细胞，这对于鉴定 NC 正常和异常发育的新标志物至关重要衍生的成体细胞类型。