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

一般程序目标和背景 * 导致牙齿形成的细胞相互作用是很好理解的，但涉及口腔粘膜牙龈的发育过程在解剖学上，组织学上和功能上与牙齿密切接触的口腔粘膜的不同部分很少受到关注。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，但他们的身份仍然是模棱两可的。有趣的是，我们和其他实验室最近的研究结果表明，牙龈结缔组织细胞亚群可能来自神经嵴（NC）和中胚层在发展过程中。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，牙龈提供了一个容易获得的来源，研究NC和中胚层来源的细胞亚群的基本特性。我们的总体长期目标是识别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，具有不同的属性。一旦鉴定和表征，这些细胞可用于寻找新的标记物，用于表征和从各种其他组织中前瞻性地分离类似的亚群。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志物的免疫反应性并通过使用NC和中胚层特异性基因报告小鼠来鉴定和定位正常牙龈中的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小鼠牙龈中分离出来，并用于蛋白质组学工具，功能测定和基因表达分析来揭示其表型，并发现新的NC和中胚层特异性细胞标记物。这些基础研究方法将首次详细鉴定和表征正常人类和小鼠牙龈中的NC和中胚层衍生细胞，并发现可用于鉴定和分离各种组织中的NC和中胚层衍生细胞的新指标。这与我们对一般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.***