Mead Timothy J
Department of Biomedical Engineering, Cleveland Clinic Lerner Research Institute, Cleveland, OH, USA.
Methods Mol Biol. 2020;2043:207-212. doi: 10.1007/978-1-4939-9698-8_17.
Understanding proteolytic remodeling of extracellular matrix involves the generation of global or conditional knockout mice by homologous recombination in embryonic stem cells or their manipulation through new advanced technologies such as CRISPR-Cas9. These models provide opportunities to understand the roles of ADAMTS genes in skeletogenesis. Whole-mount skeletal preparations are necessary for assessment of the skeletal phenotype. They allow for facile visualization of skeletal patterning, size and shape of skeletal elements, and skeletal structure. This protocol describes the staining of the murine skeleton using Alcian blue to identify cartilage and alizarin red to identify bone.
了解细胞外基质的蛋白水解重塑涉及通过胚胎干细胞中的同源重组产生全局或条件性基因敲除小鼠,或通过CRISPR-Cas9等新的先进技术对其进行操作。这些模型为了解ADAMTS基因在骨骼发育中的作用提供了机会。全骨骼标本对于评估骨骼表型是必要的。它们便于观察骨骼模式、骨骼元素的大小和形状以及骨骼结构。本方案描述了使用阿尔新蓝对小鼠骨骼进行染色以识别软骨,使用茜素红对小鼠骨骼进行染色以识别骨骼。