GENETIC EXPRESSION OF HUMAN ODONTOBLASTS

人类成牙本质细胞的基因表达

• 批准号: 6362934
• 负责人: LINDA G LEVIN
• 金额: $ 3.61万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2004-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6362934
• 关键词: dental development; gene expression; genetic library; genetic models; growth factor; human tissue; messenger RNA; model design /development; molecular cloning; northern blottings; nucleic acid sequence; odontoblasts; osteoblasts; subtraction hybridization; transcription factor; western blottings

The odontoblast is a unique cell type which is responsible for the elaboration of the organic matrix of dentin. Initially pivotal in tooth morphogenesis, it becomes equally as integral in the protection and maintenance of the fully formed tooth, Traditionally this cell has not been amenable to simplistic experimental models due to its highly differentiated and post-mitotic state. Partial characterization of the odontoblast has been accomplished using biochemical, microscopic and, most recently, molecular techniques, predominantly in rodent systems. A full characterization of the human odontoblast, however, is lacking., The overall aim of this proposal is to develop a model whereby odontoblast- specific transcripts may be identified for both quiescent and stimulated odontoblasts. We propose to do this by performing subtractive hybridization between human odontoblast mRNA and that of primary human osteoblasts. Unique transcripts will be identified by dot blot hybridization and Northern blot analysis and characterized by sequence analysis. Unique transcripts can then be used as probes in situ hybridization to verify their scope of expression in a variety of tissues. Confirmed odontoblast specific transcripts will be used as probes to screen human genomic libraries for the complete gene sequence of selected clones to study gene processing and regulation and thereby describe the role of genetic regulation in reparative dentinogenesis. Future studies would include the utilization of odontoblast-specific transcripts as differentiation markers for identifying progenitor cell pools as well as application of the proposed technique to study different activation states of human odontoblasts

成牙本质细胞是一种独特的细胞类型，负责牙本质有机基质的形成。最初在牙齿形态发生中至关重要，它在保护和维护完全形成的牙齿中同样不可或缺。传统上，由于其高度分化和有丝分裂后状态，该细胞不适合简单化的实验模型。成牙本质细胞的部分表征已经通过生物化学、显微镜和最近的分子技术完成，主要是在啮齿类动物系统中。然而，缺乏对人类成牙本质细胞的完整表征。该提案的总体目标是开发一种模型，通过该模型可以识别静止和刺激的成牙本质细胞的成牙本质细胞特异性转录本。 我们建议通过在人成牙本质细胞 mRNA 和原代人成骨细胞 mRNA 之间进行消减杂交来做到这一点。独特的转录物将通过斑点杂交和Northern印迹分析来鉴定，并通过序列分析来表征。 然后，独特的转录本可用作原位杂交探针，以验证其在各种组织中的表达范围。已确认的成牙本质细胞特异性转录本将用作探针，筛选人类基因组文库中选定克隆的完整基因序列，以研究基因加工和调控，从而描述基因调控在修复性牙本质发生中的作用。 未来的研究将包括利用成牙本质细胞特异性转录本作为识别祖细胞池的分化标记，以及应用所提出的技术来研究人类成牙本质细胞的不同激活状态