Characterization of oral mucosal neural crest and mesoderm-derived connective tissue cells

总体计划目标和背景****导致牙齿形成的细胞相互作用已经被很好地理解，但是涉及口腔粘膜牙龈发育的过程是口腔粘膜中与牙齿密切接触的解剖学、组织学和功能上不同的部分，很少受到关注。一些研究表明，牙龈结缔组织是由表型不同的成纤维细胞亚群组成的，但它们的身份仍然不明确。有趣的是，我们和其他实验室最近的发现表明，在发育过程中，牙龈结缔组织细胞亚群可能来自神经嵴（NC）和中胚层。因此，牙龈成纤维细胞亚群的表型异质性可能由其发育起源决定。我们认为，牙龈为研究NC和中胚层细胞亚群的基本特性提供了一个容易接近的来源。我们的总体长期目标是鉴定和表征牙龈中的这些成纤维细胞亚群，并利用这些信息来了解其他组织中成纤维细胞亚群的表型。*******一般假设****我们的一般假设是正常成人牙龈含有NC和中胚层来源的成纤维细胞亚群，它们具有不同的特性。一旦被鉴定和鉴定，这些细胞可以用来寻找新的标记物，用于鉴定和从各种其他组织中分离出类似的亚群。*******总体目标和方法****我们的总体长期目标是基于对不同谱系标记的免疫反应性，并通过NC和中胚层特异性基因报告小鼠，鉴定和定位正常牙龈中NC和中胚层来源的结缔组织细胞。然后，这些细胞亚群将通过免疫学和其他技术从人类和报告小鼠的牙龈中分离出来，并使用蛋白质组学工具、功能分析和基因表达分析来揭示它们的表型，并发现新的NC和中胚层特异性细胞标记。*******预期结果、意义和影响****这些基础研究方法将首次在正常人和小鼠牙龈中详细鉴定和表征NC和中胚层来源细胞，并发现可用于鉴定和分离各种组织中NC和中胚层来源细胞的新指标。这与我们对结缔组织发育过程及其发育后正常功能的理解有关***

General program goals and background****Cell interactions that lead to tooth formation are well understood but the processes involved in the development of oral mucosal gingivaan anatomically, histologically and functionally distinct part of oral mucosa that is in close contact with teethhave received little attention. Several studies have suggested that gingival connective tissue is composed of phenotypically distinct fibroblast subpopulations, but their identity has remained ambiguous. Interestingly, recent findings from our and other laboratories have indicated that gingival connective tissue cell subsets may derive from neural crest (NC) and mesoderm during development. Thus, the phenotypic heterogeneity of gingival fibroblast subpopulations may be determined by their developmental origin. We believe that gingiva provides an easily accessible source to study the fundamental properties of NC and mesoderm-derived cell subpopulations. Our general long-term goal is to identify and characterize these fibroblast subpopulations in gingiva and also use this information to understand the phenotype of fibroblast subsets in other tissues.*******General hypothesis****Our general hypothesis is that normal adult gingiva contains NC and mesoderm-derived fibroblast subpopulations that have distinct properties. Once identified and characterized, these cells can be used to find novel markers for the characterization and prospective isolation of similar subpopulations from various other tissues.*******General objectives and methods****Our general long-term goal is to identify and localize NC and mesoderm-derived connective tissue cells in normal gingiva based on immunoreactivity to distinct lineage markers and by using NC- and mesoderm-specific gene reporter mice. These cell subpopulations will then be isolated from human and reporter-mouse gingiva by immunological and other techniques and used to unravel their phenotype with proteomics tools, functional assays and gene expression analyses, and to uncover novel NC- and mesoderm-specific cell markers.*******Expected outcomes, significance and impact****These basic research approaches will identify and characterize in detail NC and mesoderm-derived cells in normal human and mouse gingiva for the first time, and uncover novel indicators that can be used to identify and isolate NC and mesoderm-derived cells in various tissues. This is relevant to our understanding of the processes involved in connective tissue development in general and its normal function post-development.***