Generation and characterization of Osterix-Cherry reporter mice.
Generation and characterization of Osterix-Cherry reporter mice.
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DOI：
10.1002/dvg.22360
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发表时间：
2013-04
期刊：
影响因子：
1.5
通讯作者：
Maye, Peter
中科院分区：
文献类型：
--
作者：
Strecker, Sara;Fu, Yu;Liu, Yaling;Maye, Peter
Osterix是一种含锌指的转录因子，是成骨细胞分化的关键调控因子。为了更好地了解Osterix在胚胎发育和成年骨骼中的时空表达，我们培育了Osterix- cherry报告小鼠。利用细菌重组技术构建了一个包含Osterix/Sp7基因的约39kb DNA片段，但不包括邻近的基因序列。Osterix报告基因的表达在胚胎、新生儿和成年小鼠中均有表征，并与骨涎蛋白(BSP)-Topaz报告基因小鼠杂交。相对于Osterix， BSP是一个更成熟的成骨细胞分化标志。与成骨细胞谱系成熟一致，在胚胎发育过程中，Osterix报告基因表达先于BSP报告基因表达，并且在空间上出现在更广泛的细胞群中。在成熟成骨细胞和骨细胞中观察到强烈的Osterix报告表达。然而，在骨髓中也观察到较弱的Osterix-Cherry阳性细胞，可能确定了早期的骨祖细胞群。对10日龄至12周龄雄性股骨组织切片中Osterix报告蛋白表达的评估显示，成骨细胞谱系中的表达持续存在，生长板中成熟软骨细胞的表达惊人地增加。此外，出生后肾脏中也短暂检测到Osterix报告基因的表达。
Osterix is a zinc finger containing transcription factor, which functions as a key regulator of osteoblast differentiation. To better understand the temporal and spatial expression of Osterix during embryonic development and in the adult skeleton, we generated Osterix-Cherry reporter mice. Bacterial recombination techniques were employed to engineer a transgenic construct, which consisted of a ~39kb DNA fragment encompassing the Osterix/Sp7 gene, but excluding adjacent gene sequences. Osterix reporter expression was characterized at embryonic, neonatal, and adult ages both by itself and in the context of a cross with Bone Sialoprotein (BSP)-Topaz reporter mice. Relative to Osterix, BSP is a more mature marker of osteoblast differentiation. In agreement with osteoblast lineage maturation, Osterix reporter expression preceded BSP reporter expression during embryonic development and spatially appeared in a much broader cell population. Strong Osterix reporter expression was observed in mature osteoblasts and osteocytes. However, weaker Osterix-Cherry positive cells were also observed in the bone marrow, possibly identifying an early osteoprogenitor cell population. Evaluation of Osterix reporter expression in male femur tissue sections from 10 days to 12 weeks of age revealed persistent expression in cells of the osteoblast lineage and a surprising increase in maturing chondrocytes of the growth plate. Also, Osterix reporter expression was transiently detected in the kidney after birth.
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