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

• 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
• 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

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.

项目摘要/摘要 神经元顶（NC）细胞的缺陷发育会引起结构性和神经系统疾病 长期对人类健康的有害影响。 NC细胞形成周围和恩的神经元和神经元 除颅面骨和软骨外，teric神经系统。 NC的早期形成和分离 来自粘附神经管的细胞是一个由细胞信号传导，表观遗传和转录物紧密调节的过程 通过改变钙粘蛋白蛋白定位，改变了细胞 - 细胞粘合剂。在结构上，作为 神经褶皱上升到在胚胎的中心相遇，以创建神经管，非神经外胚层细胞 相遇并覆盖胚胎，最终与表皮和位置区分开。神经管之后 当然，NC细胞会经历上皮到间充质转变（EMT），使其神经上皮邻居 - BOR，并在整个胚胎中迁移，侵入各种组织，并产生潜水衍生物。迅速的 转录，翻译和翻译后水平的变化允许细胞中的动态转变 极性，粘合剂和迁移。几个转录因子是编码基因的调节因子 NC细胞开发过程中的钙粘蛋白蛋白，但严重缺乏有关更快的信息 在NC EMT期间改变钙粘蛋白蛋白定位的过程。具体来说，我们旨在了解如何 微管和相关的细胞骨架因子在此发育阶段调节细胞 - 细胞粘合剂。基于 在我们发表的工作和初步数据上，在NC细胞中更新了-III微管蛋白（TUBB3）。 NC EMT。我们的目标是了解控制NC EMT和随后的复杂机制 通过定义微管元件TUBB3如何调节钙粘蛋白蛋白定位，NC分化 在NC细胞EMT和体内分化期间。此外，我们将确定TUBB3的扰动如何影响 NC EMT期间其他细胞骨架元素的结构。在AIM 1中，我们将执行收益和损失 TUBB3实验，然后进行定量分析表达和定位的变化 thelial和迁移性钙粘蛋白（CDH2，CDH1和CDH11），组织特异性标记（NC和神经管）， 细胞迁移和颅NC分化，采用转录本上的定量空间方法 鸡（Gallus Gallus）中的Tein水平，在早期阶段反映了人类发展。在AIM 2中，我们 将执行TUBB3的扰动，以确定有缺陷的TUBB3介导的微管组件 在NC EMT期间影响微管，中间丝和肌动蛋白丝的定位。和 这些实验，我们希望将传统和新颖的方法整合在一起，以理解该综合 NC EMT期间细胞粘附和细胞骨架排列之间的plex连接。我们希望贡献 我们对蛋白质网络的了解的缺失，基本要素，该蛋白质网络推动了NC的发展 细胞，这对于鉴定NC-正常和异常发育的新标记至关重要 派生的成年细胞类型。

项目成果

Effectiveness of fixation methods for wholemount immunohistochemistry across cellular compartments in chick embryos.

鸡胚胎细胞区室整体免疫组织化学固定方法的有效性。

• DOI: 10.1101/2024.03.23.586361
• 发表时间: 2024
• 期刊: bioRxiv : the preprint server for biology
• 作者: EcheverriaJr,CamiloV;Leathers,TessA;Rogers,CrystalD
• 通讯作者: Rogers,CrystalD

The graduate school guide: How to prepare for the qualifying exam and assemble a thesis/graduate committee

• DOI: 10.1002/jcp.31258
• 发表时间: 2024-04-09
• 期刊: JOURNAL OF CELLULAR PHYSIOLOGY
• 影响因子: 5.6
• 作者: Rogers，Crystal D.;Kirabo，Annet;Hinton Jr，Antentor
• 通讯作者: Hinton Jr，Antentor