Veras Matthew A, Lim Yong J, Kuljanin Miljan, Lajoie Gilles A, Urquhart Bradley L, Séguin Cheryle A
Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry The University of Western Ontario London Ontario Canada.
Bone and Joint Institute The University of Western Ontario London Ontario Canada.
JOR Spine. 2020 Jun 10;3(3):e1099. doi: 10.1002/jsp2.1099. eCollection 2020 Sep.
The comprehensiveness of data collected by "omics" modalities has demonstrated the ability to drastically transform our understanding of the molecular mechanisms of chronic, complex diseases such as musculoskeletal pathologies, how biomarkers are identified, and how therapeutic targets are developed. Standardization of protocols will enable comparisons between findings reported by multiple research groups and move the application of these technologies forward. Herein, we describe a protocol for parallel proteomic and metabolomic analysis of mouse intervertebral disc (IVD) tissues, building from the combined expertise of our collaborative team. This protocol covers dissection of murine IVD tissues, sample isolation, and data analysis for both proteomics and metabolomics applications. The protocol presented below was optimized to maximize the utility of a mouse model for "omics" applications, accounting for the challenges associated with the small starting quantity of sample due to small tissue size as well as the extracellular matrix-rich nature of the tissue.
“组学”方法收集的数据的全面性已证明，其有能力极大地改变我们对慢性复杂疾病（如肌肉骨骼疾病）分子机制的理解，以及生物标志物的识别方式和治疗靶点的开发方式。方案的标准化将使多个研究小组报告的研究结果之间能够进行比较，并推动这些技术的应用向前发展。在此，我们基于合作团队的综合专业知识，描述了一种对小鼠椎间盘（IVD）组织进行蛋白质组学和代谢组学平行分析的方案。该方案涵盖了小鼠IVD组织的解剖、样本分离以及蛋白质组学和代谢组学应用的数据分析。以下展示的方案经过优化，以最大限度地提高小鼠模型在“组学”应用中的效用，同时考虑到由于组织体积小导致起始样本量少以及组织富含细胞外基质所带来的挑战。
