Department of Craniofacial Sciences, School of Dental Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA.
Bone. 2011 Apr 1;48(4):927-37. doi: 10.1016/j.bone.2010.12.008. Epub 2010 Dec 21.
Terminal differentiation of odontoblasts from dental papilla is a long process involving several intermediate steps and changes in the transcriptional profile and expression of proteins secreted by cells in the odontoblast lineage. Transgenic mouse lines in which GFP expression is under the control of tissue- and stage specific promoters have provided powerful experimental tools for identification and isolation of cells at specific stages of differentiation along a lineage. Our previous studies showed utilization of pOBCol3.6GFP and pOBCol2.3GFP animals for identification of odontoblasts at early and late stages of polarization respectively. In the present study we used the DMP1-GFP transgenic animal as an experimental model to examine its expression during the differentiation of odontoblasts from progenitor cells in vivo and in vitro. Our observations showed that DMP1-GFP transgene is first activated in secretory/functional odontoblasts engaged in secretion of predentin and then transiently expressed at high levels in newly differentiated odontoblasts. Expression of DMP1-GFP was down-regulated in highly differentiated odontoblasts. The temporal and spatial pattern of expression of DMP1-GFP transgene closely mimics the expression of endogenous DMP1. This transgenic animal will facilitate studies of gene expression and biological functions in secretory/functional odontoblasts.
成牙本质细胞的终末分化是一个漫长的过程,涉及几个中间步骤和细胞转录谱的变化以及牙本质细胞谱系中分泌蛋白的表达。GFP 表达受组织和阶段特异性启动子控制的转基因小鼠系为鉴定和分离谱系中特定分化阶段的细胞提供了强大的实验工具。我们之前的研究表明,分别利用 pOBCol3.6GFP 和 pOBCol2.3GFP 动物鉴定极化早期和成牙本质细胞的晚期。在本研究中,我们使用 DMP1-GFP 转基因动物作为实验模型,在体内和体外检查其在成牙本质细胞从祖细胞分化过程中的表达。我们的观察结果表明,DMP1-GFP 转基因首先在参与前期牙本质分泌的分泌/功能成牙本质细胞中被激活,然后在新分化的成牙本质细胞中短暂高水平表达。在高度分化的成牙本质细胞中,DMP1-GFP 的表达被下调。DMP1-GFP 转基因的时空表达模式与内源性 DMP1 的表达模式非常相似。这种转基因动物将有助于研究分泌/功能成牙本质细胞中的基因表达和生物学功能。