Role of the novel regulator lbh in neural crest and craniofacial development

新型调节器 lbh 在神经嵴和颅面发育中的作用

• 批准号: 8647552
• 负责人: Kara E Powder
• 金额: $ 5.3万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8647552
• 关键词: Accounting; Adult; Affect; Alleles; Biological Models; Birth; CHARGE syndrome; Cellular Assay; Chondrocytes; Chondrogenesis; Cichlids; Congenital Abnormality; Craniofacial Abnormalities; Data; Defect; Development; Disease; Etiology; Evolution; Exhibits; Face; Failure; Fishes; Genes; Genomics; Goals; Heart; Homologous Gene; Human; In Situ Hybridization; Jaw; Laboratories; Lead; Length; Limb Bud; Mandible; Mediating; Messenger RNA; Methods; Micrognathism; Mitotic; Modeling; Morphology; Neural Crest; Neural Crest Cell; Pattern; Phenotype; Play; Reporter; Role; Skeleton; Specificity; Staging; Staining method; Stains; Testing; Tissues; Transcript; Up-Regulation; Variant; Zebrafish; craniofacial; innovation; knock-down; malformation; migration; mutant; novel; overexpression; premature; public health relevance; research study; skeletal

Project Summary/Abstract Craniofacial malformations are among the most common human birth defects, affecting 1/700 births. While many of the genes that underlie these defects are known, the identity and functions of many others are not. The overarching goal of this proposal is evaluate the role of a largely uncharacterized gene, limb bud and heart homolog (lbh), during zebrafish craniofacial development. This gene may mediate CHARGE syndrome, a disease of neural crest cells (NCCs) that includes craniofacial defects such as micrognathia. Additionally, we have associated this gene with the evolution of jaws in cichlids, an evolutionary model that exhibits extensive diversity in craniofacial form, many of which mimic human facial malformations. Combined, these data led to the hypothesis that lbh is a novel regulator of NCC and craniofacial development, which will be tested through the following experimental aims. Aim 1: Given the paucity of information about lbh, we will first characterize the expression pattern and cellular specificity of lbh throughout zebrafish facial development using whole-mount and sectioned in situ hybridization and comparison with known craniofacial markers. Aim 2: We will evaluate the effects of modulating lbh expression (morpholino knockdown or mRNA overexpression) on the craniofacial skeleton. Preliminary experiments indicate that depletion of Lbh results in reduction of NCCs and commensurate defects in the craniofacial skeleton. Specifically, we will determine if the cellular mechanism of this defect is due to a failure of induction, failure of migration, decreased survival, or premature differentiation of NCCs using a combination of in situ hybridization, reporter zebrafish strains, and cellular assays. Aim 3: We will analyze the functional conservation and evolution of lbh using biologically relevant alleles of lbh isolated from cichlids with differing mandible lengths. Overall, these studies will illuminate the role of lbh in craniofacial development, disease, and evolution.

项目总结/摘要 颅面畸形是人类最常见的出生缺陷之一， 影响1/700的新生儿。虽然这些缺陷背后的许多基因是 虽然我们知道，但许多其他人的身份和功能却不为人知。总体目标 这项建议的一个重要方面是评估一个很大程度上没有特征的基因，肢芽， 和心脏同源物（LBH），在斑马鱼颅面发育。该基因 可能介导CHARGE综合征，这是一种神经嵴细胞（NCC）疾病， 包括颅面缺陷如小颌畸形。此外，我们还有 将该基因与慈鲷科的颌骨进化联系起来， 在颅面形态上表现出广泛的多样性，其中许多模仿人类 面部畸形综合起来，这些数据导致了lbh是一种 NCC和颅面发育的新调节剂，将进行测试 通过以下实验目标。目标1：鉴于信息匮乏 关于lbh，我们将首先描述表达模式和细胞特异性， 在整个斑马鱼面部发育过程中使用整体安装和切片 原位杂交和与已知颅面标志物的比较。目标2：我们 将评估调节LBH表达（吗啉代敲低或 mRNA过表达）。初步实验 表明Lbh耗竭导致净捐助国减少， 颅面骨骼的缺陷具体来说，我们将确定细胞是否 这种缺陷的机制是由于诱导失败，迁移失败， 使用以下组合降低NCC的存活率或过早分化： 原位杂交、报告斑马鱼品系和细胞测定。目标3：我们 将利用生物学方法分析lbh的功能保护和进化。 从不同下颌骨长度的慈鲷中分离的lbh相关等位基因。 总之，这些研究将阐明lbh在颅面发育中的作用， 疾病和进化