Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, United States.
Department of Otolaryngology Head and Neck Surgery, Stanford University, Stanford, United States.
Elife. 2019 Nov 4;8:e50777. doi: 10.7554/eLife.50777.
Hearing and balance rely on small sensory hair cells that reside in the inner ear. To explore dynamic changes in the abundant proteins present in differentiating hair cells, we used nanoliter-scale shotgun mass spectrometry of single cells, each ~1 picoliter, from utricles of embryonic day 15 chickens. We identified unique constellations of proteins or protein groups from presumptive hair cells and from progenitor cells. The single-cell proteomes enabled the de novo reconstruction of a developmental trajectory using protein expression levels, revealing proteins that greatly increased in expression during differentiation of hair cells (e.g., OCM, CRABP1, GPX2, AK1, GSTO1) and those that decreased during differentiation (e.g., TMSB4X, AGR3). Complementary single-cell transcriptome profiling showed corresponding changes in mRNA during maturation of hair cells. Single-cell proteomics data thus can be mined to reveal features of cellular development that may be missed with transcriptomics.
听力和平衡依赖于位于内耳的小型感觉毛细胞。为了探索分化毛细胞中丰富蛋白质的动态变化，我们使用纳升级规模的单细胞shotgun 质谱法，对来自胚胎第 15 天鸡的耳石中的单个细胞（每个约 1 皮升）进行分析。我们从假定的毛细胞和祖细胞中鉴定出独特的蛋白质或蛋白质簇。单细胞蛋白质组学使我们能够使用蛋白质表达水平重新构建发育轨迹，揭示在毛细胞分化过程中表达水平大大增加的蛋白质（例如 OCM、CRABP1、GPX2、AK1、GSTO1）和那些在分化过程中减少的蛋白质（例如 TMSB4X、AGR3）。互补的单细胞转录组谱分析显示了毛细胞成熟过程中 mRNA 的相应变化。因此，单细胞蛋白质组学数据可以被挖掘出来，以揭示可能被转录组学遗漏的细胞发育特征。
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