Increasing participation of Sclerostin in postnatal bone development revealed by three-dimensional immunofluorescence morphometry.
Increasing participation of Sclerostin in postnatal bone development revealed by three-dimensional immunofluorescence morphometry.
复制标题
三维免疫荧光形态测量显示硬化蛋白越来越多地参与出生后骨骼发育。
DOI：
10.1016/j.bone.2012.06.019
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发表时间：
2012
期刊：
影响因子：
4.1
通讯作者：
Iimura T
中科院分区：
文献类型：
--
作者：
Watanabe T;Tamamura Y;Hoshino A;Makino Y;Nishimura R;Kamioka H;Yoneda T;Amagasa T;Yamaguchi A;Iimura T
共聚焦免疫荧光平铺成像显示osterix和sclerostin在3日龄、2周龄和4周龄大鼠股骨中的时空分布在组织水平上是非排他性的。进一步的三维定量免疫荧光形态计量学显示，骨细胞腔隙-小管系统中骨硬化素的分布增加，特别是在骨干中，这说明了相邻骨膜细胞中osterix和β-catenin的协同参与和耗竭。用BIO（一种GSK 3抑制剂）处理MC 3 T3-E1细胞可诱导β-catenin的稳定和osterix的核转位，并负性调节骨钙素/BGl和Dmp 1。这些结果共同表明，骨干皮质骨中硬化蛋白分布的增加似乎参与了相邻骨膜细胞中osterix和β-连环蛋白的衰减，因此可能有助于成骨细胞成熟并减少该骨部位的成骨细胞形成。我们基于共聚焦显微镜的成像分析以连贯的方式提供了sclerostin、β-catenin和osterix在组织至亚细胞水平上的时空分布的全面和详细的视图，并揭示了它们在出生后骨发育中的时空合作，从而提供了它们将成骨生长和功能性骨发育联系起来的证据。
Confocal immunofluorescence tiling imaging revealed the spatio-temporal distributions of osterix and sclerostin in femurs from 3-day-old, 2-week-old and 4-week-old rats to be reciprocally exclusive at the tissue level. Further quantitative three-dimensional immuno fluorescence morphometry demonstrated the increasing distribution of sclerostin in the osteocytic lacuno-canalicular system specifically in diaphysis, which paralleled the cooperative participation and depletion of osterix and β-catenin in adjacent periosteum cells. Treating MC3T3-E1 cells with BIO (a GSK3 inhibitor) induced the stabilization of β-catenin and nuclear translocation of osterix, and negatively regulated osteocalcin/BGLAP and Dmp1. These results collectively demonstrate that the increasing distribution of sclerostin in diaphyseal cortical bone appears to be involved in the attenuation of osterix and β-catenin in adjacent periosteum cells, thus possibly contributing to osteoblast maturation and reducing the osteoblast formation at this bone site. Our confocal microscopy-based imaging analyses provide a comprehensive and detailed view of the spatio-temporal distribution of sclerostin, β-catenin and osterix at the tissue to subcellular level in a coherent manner, and uncovered their spatio-temporal cooperation in postnatal bone development, thus providing evidence that they link skeletogenic growth and functional bone development.