The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA.
The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA.
Exp Eye Res. 2021 Jan;202:108343. doi: 10.1016/j.exer.2020.108343. Epub 2020 Nov 4.
Metabolomics is a study of the entire repertoire of metabolites in a cell at a particular time point. Here, we investigate the mouse lens at multiple embryonic and postnatal time points to establish the metabolome profile during early lens development. The lenses were isolated at six time points including embryonic day 15 (E15) and E18 and postnatal day 0 (P0), P3, P6, and P9. A total of four biological replicates of each time point, each consisting of 25 mg of lens tissue were preserved. Sample preparation was performed by protein precipitation followed by centrifugation to remove proteins and recover metabolites. The resulting extract was subjected to reverse phase/ultra-performance liquid chromatography-tandem mass spectrometry. Metabolome profiling identified a total of 353 metabolites in mouse lens, marked with an abundance of collagen, antioxidant, glycosaminoglycans, lipid, amino acid, and energy-related metabolites. A comparative metabolome analysis identified >200 metabolites exhibiting increased levels (p < 0.05) at latter time points relative to E15. Principal component analysis revealed distinct metabolomic signatures running from E15 to P9 while random forest analysis categorized lipid-, amino acid-, and nucleotide-related metabolites contributing significantly to the separation of the time points. To the best of our knowledge, this is the first report investigating the mouse lens metabolome at multiple embryonic and postnatal time points.
代谢组学是研究特定时间点细胞中所有代谢物的整体谱。在这里，我们研究了多个胚胎期和出生后的小鼠晶状体时间点，以建立早期晶状体发育过程中的代谢组特征。晶状体在六个时间点进行分离，包括胚胎第 15 天（E15）和 E18 以及出生后第 0 天（P0）、P3、P6 和 P9。每个时间点有四个生物学重复，每个重复包含 25 毫克晶状体组织。通过蛋白质沉淀进行样品制备，然后进行离心以去除蛋白质并回收代谢物。所得提取物进行反相/超高效液相色谱-串联质谱分析。代谢组学分析鉴定了小鼠晶状体中的 353 种代谢物，其中富含胶原蛋白、抗氧化剂、糖胺聚糖、脂质、氨基酸和能量相关代谢物。比较代谢组分析鉴定出 200 多种代谢物在后一时间点的水平增加（p < 0.05）相对 E15。主成分分析显示从 E15 到 P9 存在明显的代谢组特征，而随机森林分析将脂质、氨基酸和核苷酸相关代谢物分类为对时间点分离有重要贡献的代谢物。据我们所知，这是首次在多个胚胎期和出生后时间点研究小鼠晶状体的代谢组学。
