Dentin matrix protein 1 expression during osteoblastic differentiation, generation of an osteocyte GFP-transgene
Dentin matrix protein 1 expression during osteoblastic differentiation, generation of an osteocyte GFP-transgene
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DOI：
10.1016/j.bone.2004.03.006
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发表时间：
2004-07-01
期刊：
影响因子：
4.1
通讯作者：
Rowe, DW
中科院分区：
文献类型：
--
作者：
Kalajzic, I;Braut, A;Rowe, DW
我们以前的研究已经证明，启动子-绿色荧光蛋白（GFP）转基因可用于鉴定和分离活的原代骨细胞培养物中的前成骨细胞阶段（pOBCol3.6GFP）和成熟成骨细胞阶段（pOBCol2.3GFP）的细胞群。这种策略形成了欣赏谱系的细胞异质性和将基因功能与细胞分化相关联的基础。这种方法的缺点是缺乏矿化基质中晚期成骨细胞和成熟骨细胞的选择性标记。在这项研究中，我们已经研究了在小鼠骨髓基质和颅骨成骨细胞培养，并在骨，和颅骨在体内的表达。此外，我们已经产生了转基因小鼠，利用小鼠DMP 1顺式调节系统驱动GFP作为活骨细胞的标记。转基因表达针对矿化组织，并与内源基因的表达高度相关。在股骨和颅骨的组织切片和原代细胞培养物中观察到骨细胞限制性GFP表达。这种转基因模型的建立将有助于研究终末分化骨细胞的基因表达和生物学功能。(C)2004年爱思唯尔公司All rights reserved.
Our previous studies have demonstrated that promoter-green fluorescent protein (GFP) transgenes can be used to identify and isolate populations of cells at the preosteoblastic stage (pOBCol3.6GFP) and at the mature osteoblastic stage (pOBCol2.3GFP) in living primary bone cell cultures. This strategy forms the basis for appreciating the cellular heterogeneity of lineage and relating gene function to cell differentiation. A weakness of this approach was the lack of a selective marker for late ostcoblasts and mature osteocytes in the mineralized matrix. In this study, we have examined the expression of DMP-1 mRNA in murine marrow stromal and calvarial osteoblast cultures, and in bone, and calvaria in vivo. Furthermore, we have generated transgenic mice utilizing a mouse DMP1 cis-regulatory system to drive GFP as a marker for living osteocytes. Transgene expression was directed to mineralized tissues and showed a high correlation with the expression of the endogenous gene. Osteocyte-restricted expression of GFP was observed in histological sections of femur and calvaria and in primary cell cultures. Generation of this transgenic model will facilitate studies of gene expression and biological functions in these terminally differentiated bone cells. (C) 2004 Elsevier Inc. All rights reserved.