Matsuzawa M, Sheu T-J, Lee Y-J, Chen M, Li T-F, Huang C T, Holz J D, Puzas J E
Department of Orthopaedics, Center for Musculoskeletal Research, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, USA.
J Periodontal Res. 2009 Jun;44(3):289-96. doi: 10.1111/j.1600-0765.2008.01091.x.
While it has long been known that amelogenin is essential for the proper development of enamel, its role has generally been seen as structural in nature. However, our new data implicate this protein in the regulation of cell signaling pathways in periodontal ligament cells and osteoblasts. In this article we report the successful purification of a recombinant mouse amelogenin protein and demonstrate that it has signaling activity in isolated mouse calvarial cells and human periodontal ligament cells.
To determine the regulatory function of canonical Wnt signaling by amelogenin, we used TOPGAL transgenic mice. These mice express a beta-galactosidase transgene under the control of a LEF/TCF and beta-catenin-inducible promoter. To investigate in greater detail the molecular mechanisms involved in the beta-catenin signaling pathway, isolated osteoblasts and periodontal ligament cells were exposed to full-length recombinant mouse amelogenin and were evaluated for phenotypic changes and beta-catenin signaling using a TOPFLASH construct and the LacZ reporter gene.
In these in vitro models, we showed that amelogenin can activate beta-catenin signaling.
Using the TOPGAL transgenic mouse we showed that amelogenin expression in vivo is localized mainly around the root, the periodontal ligament and the alveolar bone.
长期以来人们都知道釉原蛋白对牙釉质的正常发育至关重要,其作用通常被认为本质上是结构性的。然而,我们的新数据表明该蛋白参与牙周膜细胞和成骨细胞中细胞信号通路的调节。在本文中,我们报告了重组小鼠釉原蛋白的成功纯化,并证明其在分离的小鼠颅骨细胞和人牙周膜细胞中具有信号活性。
为了确定釉原蛋白对经典Wnt信号的调节功能,我们使用了TOPGAL转基因小鼠。这些小鼠在LEF/TCF和β-连环蛋白诱导型启动子的控制下表达β-半乳糖苷酶转基因。为了更详细地研究β-连环蛋白信号通路涉及的分子机制,将分离的成骨细胞和牙周膜细胞暴露于全长重组小鼠釉原蛋白,并使用TOPFLASH构建体和LacZ报告基因评估其表型变化和β-连环蛋白信号。
在这些体外模型中,我们表明釉原蛋白可以激活β-连环蛋白信号。
使用TOPGAL转基因小鼠,我们表明体内釉原蛋白的表达主要定位于牙根、牙周膜和牙槽骨周围。