Neural crest and placode cell interactions during cranial gangliogenesis

颅神经节发生过程中神经嵴和基板细胞的相互作用

• 批准号: 8928593
• 负责人: LISA A TANEYHILL
• 金额: $ 37.64万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-18 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8928593
• 关键词: Address; Adherens Junction; Adhesions; Behavior; Binding; Biochemical; Biochemistry; Biological; Biological Assay; Cadherins; Candidate Disease Gene; Cell Communication; Cells; Cellular Structures; Cellular biology; Cephalic; Chick Embryo; Complex; Data; Development; Disease; Embryo; Embryology; Embryonic Development; Environment; Face; Filopodia; Ganglia; Gap Junctions; Gene Expression Profile; Goals; Growth; Health; Heart; Heart Part; Human; Human Development; Human body; Image; Immunohistochemistry; In Situ Hybridization; In Vitro; Inherited; Knowledge; Lead; Life; Light; Malignant Neoplasms; Mediating; Methods; Mission; Modeling; Molecular; Molecular Biology; Molecular Profiling; Mus; N-Cadherin; Nature; Nerve; Neural Crest; Neural Crest Cell; Organ; Pathway interactions; Pattern; Peripheral Nervous System; Pigments; Play; Population; Process; Protein Isoforms; Proteins; Public Health; Publishing; Quality of life; RNA Sequences; Reporting; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Sensory; Skeleton; Skin; Staging; Structure; Structure of trigeminal ganglion; Syndrome; Testing; Time; Tissues; Work; base; bone; cadherin 7; cell motility; cell type; cellular imaging; craniofacial; genetic manipulation; human disease; improved; in vivo; in vivo Model; in vivo imaging; innovation; malformation; migration; novel; relating to nervous system; repaired; transcriptome sequencing

DESCRIPTION (provided by applicant): Cranial ganglia of the peripheral nervous system are critical for integrating sensory information and controlling cell movements. The appropriate coalescence of both neural crest cells (NCCs) and placode cells (PCs) is required for the proper formation of many of these ganglia. Migratory NCCs also differentiate to create other cell types, including the craniofacial skeleton and skin pigment cells. Abnormalities that occur during NCC and PC development are thus directly responsible for many human congenital and hereditary malformations, diseases and cancers. Cranial ganglia assembly involves the creation of new junctions between NCCs and PCs, facilitating their interactions with one another and with their surroundings, but the molecular composition of these junctional complexes is not known. Our published and preliminary data indicate that NCCs and PCs each express a different repertoire of cadherins and catenins, which may function to mediate both NCC and PC migration as well as the formation of novel heterophilic adherens junctions between NCCs and PCs during cranial ganglia assembly. Such heterophilic interactions have not been reported between different cell types, and, together with data on other junctions, could prove to be paradigm-shifting in defining how different cells coalesce to form multi-cellular structures. Moreover, we report for the first time the effects of perturbation of gap junctions, which play crucial roles in cell-cell communication, on chick NCC migration. In light of our data, we hypothesize that cranial ganglia assembly is dependent upon the temporal formation of heterophilic cadherin-based adherens junctions between migratory NCCs and PCs, followed by the creation of other anchoring and communicating junctions. The Specific Aims of this application are to: 1) determine the requirement for cadherins and catenins in mediating NCC and PC migration, 2) define the role of cadherins and catenins in cranial ganglia formation, and 3) define additional junctional complexes that orchestrate cranial gangliogenesis. In Aim 1, we will perturb NCC and PC cadherins and catenins by adapting the use of photo morpholinos and inducible expression constructs to the chick embryo and documenting changes in NCC and PC migration through in vitro and in vivo imaging assays. In Aim 2, we will use the above perturbation methods and biochemical assays we have pioneered in the chick embryo to evaluate the function of cadherins and catenins, and heterophilic interactions, in cranial ganglia assembly. In Aim 3, we will perform RNA-sequencing of chick trigeminal ganglia to elucidate a molecular signature for gangliogenesis, facilitating the identification of other junctional complexes that mediate NCC-PC interactions. The proposed research is innovative because it takes a multi-disciplinary approach that combines embryology, biochemistry, and cell and molecular biology to examine how different cell populations interact during tissue formation. These studies are significant because the results will enhance our understanding of the mechanisms underlying the intercellular interactions required to create tissues and organs and could lead to the development of new treatments for human diseases.

描述（由申请人提供）：周围神经系统的颅神经节对于整合感觉信息和控制细胞运动至关重要。许多神经节的正确形成需要神经嵴细胞（NCC）和基板细胞（PC）的适当聚结。迁移性 NCC 还会分化产生其他细胞类型，包括颅面骨骼和皮肤色素细胞。因此，NCC 和 PC 发育过程中发生的异常直接导致许多人类先天性和遗传性畸形、疾病和癌症。颅神经节组装涉及 NCC 和 PC 之间新连接的创建，促进它们彼此之间以及与周围环境的相互作用，但这些连接复合体的分子组成尚不清楚。我们发表的初步数据表明，NCC 和 PC 各自表达不同的钙粘蛋白和连环蛋白，这可能起到介导 NCC 和 PC 迁移以及在颅神经节组装过程中 NCC 和 PC 之间新型异嗜粘附连接的形成的作用。不同细胞类型之间的这种异嗜性相互作用尚未被报道，并且与其他连接的数据一起，可能被证明在定义不同细胞如何结合形成多细胞结构方面具有范式转变。此外，我们首次报告了间隙连接的扰动对雏鸡 NCC 迁移的影响，间隙连接在细胞间通讯中发挥着至关重要的作用。根据我们的数据，我们假设颅神经节组装依赖于迁移性 NCC 和 PC 之间基于异嗜钙粘蛋白的粘附连接的时间形成，随后创建其他锚定和通信连接。该申请的具体目标是：1) 确定钙粘蛋白和连环蛋白在介导 NCC 和 PC 迁移中的需求，2) 定义钙粘蛋白和连环蛋白在颅神经节形成中的作用，以及 3) 定义协调颅神经节生成的其他连接复合物。在目标 1 中，我们将通过在鸡胚中使用光吗啉和诱导表达构建体来扰乱 NCC 和 PC 钙粘蛋白和连环蛋白，并通过体外和体内成像测定记录 NCC 和 PC 迁移的变化。在目标 2 中，我们将使用我们在鸡胚胎中首创的上述扰动方法和生化测定来评估颅神经节组装中钙粘蛋白和连环蛋白的功能以及异嗜性相互作用。在目标 3 中，我们将对鸡三叉神经节进行 RNA 测序，以阐明神经节发生的分子特征，从而促进介导 NCC-PC 相互作用的其他连接复合物的识别。拟议的研究具有创新性，因为它采用了结合胚胎学、生物化学以及细胞和分子生物学的多学科方法来研究不同细胞群在组织形成过程中如何相互作用。这些研究意义重大，因为结果将增强我们对创造组织和器官所需的细胞间相互作用的机制的理解，并可能导致人类疾病新疗法的开发。