Department of Molecular Genetics, University of Toronto, Toronto, Canada.
J Bone Miner Res. 2014 Jun;29(6):1412-23. doi: 10.1002/jbmr.2177.
By using a genome-wide N-ethyl-N-nitrosourea (ENU)-induced dominant mutagenesis screen in mice, a founder with low bone mineral density (BMD) was identified. Mapping and sequencing revealed a T to C transition in a splice donor of the collagen alpha1 type I (Col1a1) gene, resulting in the skipping of exon 9 and a predicted 18-amino acid deletion within the N-terminal region of the triple helical domain of Col1a1. Col1a1(Jrt) /+ mice were smaller in size, had lower BMD associated with decreased bone volume/tissue volume (BV/TV) and reduced trabecular number, and furthermore exhibited mechanically weak, brittle, fracture-prone bones, a hallmark of osteogenesis imperfecta (OI). Several markers of osteoblast differentiation were upregulated in mutant bone, and histomorphometry showed that the proportion of trabecular bone surfaces covered by activated osteoblasts (Ob.S/BS and N.Ob/BS) was elevated, but bone surfaces undergoing resorption (Oc.S/BS and N.Oc/BS) were not. The number of bone marrow stromal osteoprogenitors (CFU-ALP) was unaffected, but mineralization was decreased in cultures from young Col1a1(Jrt) /+ versus +/+ mice. Total collagen and type I collagen content of matrices deposited by Col1a1(Jrt) /+ dermal fibroblasts in culture was ∼40% and 30%, respectively, that of +/+ cells, suggesting that mutant collagen chains exerted a dominant negative effect on type I collagen biosynthesis. Mutant collagen fibrils were also markedly smaller in diameter than +/+ fibrils in bone, tendon, and extracellular matrices deposited by dermal fibroblasts in vitro. Col1a1(Jrt) /+ mice also exhibited traits associated with Ehlers-Danlos syndrome (EDS): Their skin had reduced tensile properties, tail tendon appeared more frayed, and a third of the young adult mice had noticeable curvature of the spine. Col1a1(Jrt) /+ is the first reported model of combined OI/EDS and will be useful for exploring aspects of OI and EDS pathophysiology and treatment.
通过在小鼠中进行全基因组 N-乙基-N-亚硝脲(ENU)诱导显性突变筛选,鉴定出一个具有低骨密度(BMD)的突变体。图谱绘制和测序显示,Ⅰ型胶原蛋白(Col1a1)基因的一个剪接供体位点发生 T 到 C 的转换,导致外显子 9 跳跃,以及 Col1a1 三螺旋结构域的 N 端区域预测的 18 个氨基酸缺失。Col1a1(Jrt)/+ 小鼠体型较小,BMD 降低,与骨体积/组织体积(BV/TV)减少和小梁数量减少有关,此外还表现出机械强度弱、易碎、易骨折的骨骼,这是成骨不全症(OI)的标志。突变骨中的几种成骨细胞分化标志物上调,组织形态计量学显示,被激活的成骨细胞(Ob.S/BS 和 N.Ob/BS)覆盖的小梁骨表面比例升高,但正在吸收的骨表面(Oc.S/BS 和 N.Oc/BS)没有升高。骨髓基质成骨细胞前体(CFU-ALP)数量没有受到影响,但来自年轻 Col1a1(Jrt)/+ 与 +/+ 小鼠的培养物中的矿化减少。Col1a1(Jrt)/+ 真皮成纤维细胞培养物中沉积的基质总胶原和 I 型胶原含量分别约为 +/+ 细胞的 40%和 30%,表明突变胶原链对 I 型胶原生物合成具有显性负效应。突变胶原纤维在骨骼、肌腱和真皮成纤维细胞体外沉积的细胞外基质中的直径也明显小于 +/+ 纤维。Col1a1(Jrt)/+ 小鼠还表现出与埃勒斯-当洛斯综合征(EDS)相关的特征:它们的皮肤拉伸性能降低,尾巴肌腱显得更磨损,三分之一的成年小鼠脊柱明显弯曲。Col1a1(Jrt)/+ 是报道的首个 OI/EDS 联合模型,将有助于探索 OI 和 EDS 病理生理学和治疗的各个方面。