Endocrine Research Unit, Veterans Administration Medical Center, University of California San Francisco, San Francisco, CA 94121, USA.
J Bone Miner Res. 2011 Apr;26(4):822-32. doi: 10.1002/jbmr.273.
Osteoblasts play a critical role in the maintenance of bone mass through bone formation and regulation of bone resorption. Targeted expression of a constitutively active engineered G(i)-coupled G protein-coupled receptor (GPCR) to osteoblasts in vivo leads to severe osteopenia. However, little is known about the role of endogenous receptor-mediated G(i) signaling in regulating osteoblast function. In this study, we investigated the skeletal effects of blocking G(i)-coupled signaling in osteoblasts in vivo. This was accomplished by transgenic expression of the catalytic subunit of pertussis toxin (PTX) under control of the collagen Iα 2.3-kb promoter. These mice, designated Col1(2.3)(+)/PTX(+), showed increased cortical thickness at the femoral midshaft at 12 weeks of age. This correlated with increased periosteal bone formation associated with expanded mineralizing surface observed in 8-week-old mice of both genders. The cancellous bone phenotype of the Col1(2.3)(+)/PTX(+) mice was sexually dimorphic, with increases in fractional bone volume at the distal femur seen only in females. Similarly, while cancellous bone-formation rates were unchanged in males, they could not be quantified for female Col1(2.3)(+)/PTX(+) mice owing to the disorganized nature of the labeling pattern, which was consistent with rapid formation of woven bone. Alterations in osteoclast activity did not appear to participate in the phenotype. These data demonstrate that G(i)-coupled signaling by GPCRs endogenous to osteoblasts plays a complex role in the regulation of bone formation in a manner that is dependent on both gender and the anatomic site within bone.
成骨细胞通过骨形成和骨吸收调节在维持骨量方面起着关键作用。在体内将组成型激活的工程化 G(i)偶联 G 蛋白偶联受体 (GPCR) 靶向表达于成骨细胞会导致严重的骨质疏松症。然而,对于内源性受体介导的 G(i)信号在调节成骨细胞功能中的作用知之甚少。在这项研究中,我们研究了体内阻断成骨细胞中 G(i)偶联信号对骨骼的影响。这是通过在 I 型胶原 2.3-kb 启动子的控制下表达百日咳毒素 (PTX) 的催化亚单位来实现的。这些命名为 Col1(2.3)(+)/PTX(+)的小鼠在 12 周龄时股骨中段的皮质厚度增加。这与在 8 周龄的雌雄小鼠中观察到的与扩大的矿化表面相关的骨膜骨形成增加有关。Col1(2.3)(+)/PTX(+)小鼠的松质骨表型具有性别二态性,仅在雌性中观察到远端股骨的骨体积分数增加。同样,虽然雄性的骨小梁形成率没有变化,但由于标记模式的紊乱性质,无法对雌性 Col1(2.3)(+)/PTX(+)小鼠进行定量,这与编织骨的快速形成一致。破骨细胞活性的改变似乎没有参与表型。这些数据表明,成骨细胞内源性 GPCR 的 G(i)偶联信号在调节骨形成方面发挥了复杂的作用,这种作用依赖于性别和骨骼内的解剖部位。