Zebrafish Jaw Morphogenesis: Understanding Development Beyond the Embryo

描述(申请人提供)：斑马鱼已迅速成为研究早期颅面发育和图案的重要模型。这一系统的工作对指导细胞活动和组织相互作用的无数遗传信号提供了重要的见解，这些信号是正确组装咽部骨骼所必需的。不幸的是，我们对胚胎期以外的颅面发育的了解还远远不够完整。在这里，我们建议利用实验上易驯化的斑马鱼来研究胚胎后颅面的发育、生长和重塑。提出了三个具体目标。首先，我们将使用复杂的定量形状分析来表征两个野生型斑马鱼斑点的头面部生长。这一目标的目的是定义斑马鱼正常颅面生长和重塑的解剖学变化，并评估遗传背景对这些过程的影响程度。接下来，我们将使用相同的统计方法来分析在成纤维细胞生长因子缺乏的情况下的颅面生长。成纤维细胞生长因子是调节骨沉积和骨吸收的细胞间信号分子。我们将使用热休克诱导的显性阴性的转基因斑马鱼的成纤维细胞生长因子受体系来干扰成纤维细胞生长因子的活性，并检测其对颅面生长和重塑的影响。最后，在与塔夫茨牙医学院的帕梅拉·C·耶利克合作进行的一项正在进行的筛查中，将对新发现的矿化组织斑马鱼突变体进行胚胎后颅面发育分析。这一目标的目的是鉴定和鉴定在骨骼生长和重塑过程中表现出阶段性缺陷的新的突变体，并开始阐明调节这些过程的信号通路。这些数据将为我们理解颅面发育、生长和重塑的协调过程做出新的贡献。公共卫生相关性：这项建议试图通过将实验胚胎学与统计形状分析相结合，在一个重要的模型系统--斑马鱼--中了解胚胎后颅面发育。

DESCRIPTION (provided by applicant): The zebrafish has quickly emerged as an important model for the study of early craniofacial developmental and patterning. Work in this system has provided significant insights into the myriad genetic signals that direct the cellular activities and tissue interactions necessary for proper assembly of the pharyngeal skeleton. Unfortunately our understanding of craniofacial development beyond embryonic stages is far less complete. Here we propose to exploit the experimentally tractable zebrafish to investigate post-embryonic craniofacial development, growth, and remodeling. Three specific aims are proposed. First, we will characterize craniofacial growth in two wild-type zebrafish stains using sophisticated quantitative shape analyses. The goal of this aim is to define the anatomical shifts that characterize normal craniofacial growth and remodeling in zebrafish, and to assess the degree to which genetic background influences these processes. Next, we will use the same statistical methods to analyze craniofacial growth in the context of Fgf-deficiency. Fgf's are intercellular signaling molecules that regulate bone deposition and resorption. We will use a heat-shock inducible dominent- negative Fgf receptor line of transgenic zebrafish to disrupt Fgf activity and examine the effects on craniofacial growth and remodeling. Finally, post-embryonic craniofacial development will be analyzed in newly identified mineralized tissue zebrafish mutants in an ongoing screen being performed in collaboration with Pamela C. Yelick at Tufts School of Dental Medicine. The goal of this aim is to identify and characterize novel mutants that exhibit stage-specific defects in skeletal growth and remodeling, and to begin to elucidate the signaling pathways that regulate these processes. These data will make novel contributions to our understanding of the coordinated processes of craniofacial development, growth and remodeling. PUBLIC HEALTH RELEVANCE: This proposal seeks to understand post-embryonic craniofacial development in an important model system - the zebrafish - by combining experimental embryology with statistical shape analyses.