The Extreme Anterior Domain and Face Formation

极端前域和面部形成

基本信息

批准号：
9302725
负责人：
Hazel L Sive
金额：
$ 43.88万
依托单位：
WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-03-15 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9302725
关键词：
Address Animal Cap Anterior Anterior Pituitary Gland Anterior nares Biological Assay Birth Bradykinin Cells Cephalic Complement Cytoskeleton Data Deformity Development Dose Ectoderm Embryo Endoderm Excision Face Frontonasal Prominence Funding Gene Targeting Genes Human Immunoglobulin G Income Individual Kininogenase Kinins Life Mediating Modeling Muscle Nanotubes Neural Crest Neural Crest Cell Nitric Oxide Oral cavity Outcome Pathway interactions Peptides Prenatal Diagnosis Production Proteins Rana Reporter Role S-nitro-N-acetylpenicillamine Signal Pathway Signal Transduction Site Surface Ectoderm Testing Tissues Xenopus laevis beta catenin bone convergent extension craniofacial implantation improved in vivo imaging inhibitor/antagonist insight loss of function migration mutant novel psychologic public health relevance response sensor

项目摘要

DESCRIPTION (provided by applicant): This proposal will define signaling mechanisms associated with the extreme anterior domain (EAD) and face formation using the frogs Xenopus laevis and X. tropicalis as models. The EAD is a conserved embryonic region where ectoderm and endoderm are juxtaposed, that develops into the mouth, anterior pituitary and nostrils. In the previous funding period, we made several novel findings. (1) The EAD is an organizing center necessary for cranial neural crest (CNC) development, using the Kinin-Kallikrein pathway and nitric oxide (NO); (2) Incoming CNC induces the EAD to undergo convergent extension, a novel step in mouth formation, using the Wnt/PCP pathway; (3) Wnt antagonists frzb1+crescent that are localized in the EAD act globally, in other facial regions. Our data give insight into the earliest stages of facial development and suggest two hypotheses with high impact that will be addressed in this proposal. Wnt antagonists from the EAD regulate frontonasal prominence (FNP) and first arch CNC development. The EAD signals through Bradykinin and nitric oxide to regulate CNC migration, in a concentration-dependent manner. There are two Aims. The first Aim will delineate the role of Frzb1+Crescent derived from the EAD in neural crest development. The role(s) of EAD-derived Frzb1+Crescent in FNP and first arch CNC determination, migration, proliferation and survival will be assessed after local loss of function (LOF). X. tropicalis mutants will complement and extend analyses. Frzb1+Crescent will be assayed for sufficiency to direct neural crest development, using heterologous cells expressing Frzb1 or beads soaked in Frzb-IgG protein. Ability of Frzb1+Crescent to inhibit β-catenin-mediated Wnt reporter activity in the FNP and first arch CNC will be assessed. Signaling pathways and candidate target genes modulated by frzb1+crescent will be determined. The second Aim will define the role of the EAD and the Kinin-Kallikrein pathway in modulating nitric oxide (NO) signaling and CNC migration. Cells in the developing facial region which produce NO will be identified, using the NO sensor DAF2 and NO-sensor nanotubes. A requirement for the EAD and EAD Kinin-Kallikrein factors in facial NO production will be assessed by extirpation and local LOF. cCNC cells will be assayed for ability to respond directly to Bdk peptides or NO in the embryo or in culture. Downstream effects of NO on target cells will be examined focusing on changes in signaling pathways and the cytoskeleton. The face is the defining feature of the individual human. Facial abnormalities are frequent (~1/700 births), resulting in physical and psychological disturbances. Much of the face may be impacted by reduced activity of the EAD organizer. The data obtained here will inform pre-natal diagnosis and correction, and are highly significant in the craniofacial field.

描述（由申请人提供）：该提案将使用非洲爪蟾和热带爪蟾作为模型来定义与极端前部区域（EAD）和面部形成相关的信号传导机制。EAD是一个保守的胚胎区域，其中外胚层和内胚层并置。，发育成口腔、垂体前叶和鼻孔。在之前的资助期间，我们取得了一些新的发现（1）EAD 是一个必要的组织中心。颅神经嵴 (CNC) 发育，利用激肽-激肽释放酶途径和一氧化氮 (NO)；(2) 传入的 CNC 诱导 EAD 进行会聚伸展，这是利用 Wnt/PCP 途径形成口腔的新步骤 (3) ) 位于 EAD 中的 Wnt 拮抗剂 frzb1+crescent 在全球范围内发挥作用，我们的数据可以深入了解其他面部区域。面部发育的最早阶段，并提出了两个具有重大影响的假设，EAD 的 Wnt 拮抗剂调节额鼻突出 (FNP) 和第一弓 CNC 发育，EAD 通过缓激肽和一氧化氮发出信号来调节 CNC 迁移，有两个目标。第一个目标将描述源自 EAD 的 Frzb1+Crescent 在神经嵴发育中的作用。 FNP 和第一弓 CNC 测定中的 EAD 衍生的 Frzb1+Crescent 将在局部功能丧失 (LOF) 后进行评估，并对 Frzb1+Crescent 进行充分分析。直接神经嵴发育，使用表达 Frzb1 的异源细胞或浸泡在 Frzb-IgG 蛋白中的珠子抑制 Frzb1+Crescent 的能力。将评估 FNP 和第一弓 CNC 中 β-连环蛋白介导的 Wnt 报告基因活性，并确定由 frzb1+crescent 调节的候选靶基因。第二个目标将确定 EAD 和激肽激肽释放酶通路的作用。使用 NO 传感器 DAF2 和 NO 传感器纳米管，可以识别正在发育的面部区域中产生 NO 的细胞。面部 NO 产生中的 EAD 和 EAD 激肽激肽释放酶因子将通过切除进行评估，并且将检测局部 LOF 细胞直接响应胚胎或培养物中的 Bdk 肽或 NO 的能力。将检查细胞，重点关注信号通路和细胞骨架的变化。面部是人类个体的决定性特征。面部异常很常见（约 1/700 的出生），从而导致身体缺陷。面部的大部分区域可能会受到 EAD 组织者活动减少的影响。此处获得的数据将为产前诊断和矫正提供信息，并且在颅面领域非常重要。