Molecular Interactions During Neural Crest Formation

神经嵴形成过程中的分子相互作用

• 批准号: 7934225
• 负责人: MARTIN I. GARCIA-CASTRO
• 金额: $ 10万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-08 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7934225
• 关键词: Address; Back; Binding; Biological Assay; Cartilage; Cells; Cephalic; Cleft Palate; Collagen; Congenital Heart Defects; Defect; Development; Developmental Biology; Diagnosis; Embryo; Embryology; Environment; Enzymes; Epiblast; Event; Fibroblast Growth Factor; Future; Gastrula; Health; Heart; Human; In Vitro; Inherited; Lead; Light; Location; Maps; Modification; Molecular; Molecular Biology; Neural Crest; Neural Crest Cell; Neural Fold; Neuroglia; Neurons; Peripheral Nervous System; Play; Proteins; Reagent; Research; Risk; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Specific qualifier value; Staging; Stem cells; System; Testing; Time; Tissues; To specify; Ubiquitin Like Proteins; WNT Signaling Pathway; Yeasts; blastocyst; blastomere structure; bone; cartilage cell; combinatorial; craniofacial; flexibility; gastrulation; in vivo; melanocyte; melanoma; molecular marker; prospective; protein structure; relating to nervous system; research study; transcription factor; tumor; yeast two hybrid system

DESCRIPTION (provided by applicant): Neural crest stem cells are involved in human illnesses including tumors and craniofacial and heart malformations. They generate most of the cells of the peripheral nervous system, craniofacial bone and cartilage, cells of the outflow tract of the heart and melanocytes amongst other derivatives. Studying the basic developmental biology of the neural crest is a critical step to better understand, diagnose, and treat these human conditions. The long term objective of this proposal is to study the molecular interactions at play during neural crest cell formation at the earliest possible stages. Therefore it is imperative to identify the time, place and cells or tissues involved in their induction. It has been recently shown that a region of the epiblast is specified to form neural crest cells during gastrulation, these same region will express Pax7 soon after, and that Pax7 is required for neural crest formation. In light of these findings it is proposed to investigate when, where and how neural crest cells are induced and specified though a combination of classic embryology and molecular biology approaches. The specific aims are: 1) identify time, location and tissues involved in the induction and specification of the neural crest; 2) characterize the participation of BMP, FGF and WNT molecules in neural crest formation; and 3) Characterize Pax7 molecular interactions during neural crest formation. These aims are guided by a set of testable hypotheses that analyze neural crest cell development at the blastula/gastrula stage, challenge the accepted cannon of their induction through neural and non-neural/mesodermal interactions, and analyze at the molecular level the participation of three signaling pathways and the mode of action of Pax7. In vivo and in vitro experiments will record the progression of neural crest developments with a battery of molecular markers. To this end, specification and induction maps will be obtained through collagen explants and grafts to naive embryonic environments from epiblast tissues to be tested (specific aim 1); distinct BMP, FGF and WNT signaling conditions (including combinatorial activation and inhibition approaches, along with specific cell autonomous and non-cell autonomous reagents), will be applied to prospective and non-prospective neural crest tissues before and after induction (specific aim 2); and finally we will characterize the interaction between Pax7 and Ubc9, and their role during neural crest development, (specific aim 3).

描述(申请人提供)：神经脊干细胞与人类疾病有关，包括肿瘤、颅面和心脏畸形。它们产生周围神经系统的大部分细胞，颅面骨和软骨，心脏流出道细胞和黑素细胞以及其他衍生细胞。研究神经脊的基本发育生物学是更好地了解、诊断和治疗这些人类疾病的关键一步。这项建议的长期目标是研究在神经脊细胞形成的早期阶段所起作用的分子相互作用。因此，必须确定它们诱导的时间、地点和细胞或组织。最近的研究表明，在原肠胚形成过程中，外胚层的一个区域被指定为形成神经脊细胞，这些区域很快就会表达Pax7，而Pax7是形成神经脊所必需的。根据这些发现，建议通过结合经典胚胎学和分子生物学的方法来研究神经脊细胞何时、何地以及如何被诱导和指定。其具体目的是：1)确定神经脊的诱导和规范所涉及的时间、位置和组织；2)表征BMP、成纤维细胞生长因子和WNT分子在神经脊形成中的作用；以及3)表征神经脊形成过程中的Pax7分子相互作用。这些目标是由一系列可检验的假说指导的，这些假说分析了胚泡/原肠胚期神经脊细胞的发育，挑战了公认的通过神经和非神经/中胚层相互作用诱导它们的大炮，并在分子水平上分析了三条信号通路的参与和Pax7的作用方式。体内和体外实验将使用一组分子标记来记录神经脊发育的进展。为此，将通过胶原外植体和移植到待测试的上胚层组织的幼稚胚胎环境中获得规范和诱导图(特定目标1)；不同的BMP、成纤维细胞生长因子和WNT信号条件(包括组合激活和抑制方法，以及特定的细胞自主和非细胞自主试剂)将应用于诱导前后的预期和非预期的神经脊组织(特定目标2)；最后，我们将表征Pax7和Ubc9之间的相互作用，以及它们在神经脊发育过程中的作用(特定目标3)。