Standardized Profiling of The Membrane-Enriched Proteome of Mouse Dorsal Root Ganglia (DRG) Provides Novel Insights Into Chronic Pain
Standardized Profiling of The Membrane-Enriched Proteome of Mouse Dorsal Root Ganglia (DRG) Provides Novel Insights Into Chronic Pain
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DOI：
10.1074/mcp.m116.058966
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发表时间：
2016-06-01
影响因子：
7
通讯作者：
Schmidt, Manuela
中科院分区：
文献类型：
--
作者：
Rouwette, Tom;Sondermann, Julia;Schmidt, Manuela
慢性疼痛是一种复杂的疾病，治疗选择有限。为了剖析其分子基础，已经进行了几项分析工作。然而，生成的结果往往是不一致和不重叠的，这在很大程度上是由于固有的技术限制。新兴的数据独立获取(DIA)-质谱学(MS)有可能提供无偏见、可重现和定量的蛋白质组图谱--这是实验标准化的先决条件。在这里，我们设计了一种基于DIA的蛋白质组学工作流程，以描述两种慢性疼痛(炎症和神经病理性)小鼠模型中背根神经节(DRG)蛋白丰度的变化。我们生成了一个包含3067个DRG蛋白的DRG特异性谱库，这使得在任何实验室都可以通过DIA-MS对其进行标准化定量。利用这个资源，我们以前所未有的重复性在慢性疼痛模型和各自的对照的每个生物复制中分析了2526个DRG蛋白。我们检测到了许多差异调节蛋白，其中大部分显示出疼痛模型特异性。我们的方法概括了已知的生物学，并发现了数十种以前在体感系统中尚未被表征的蛋白质。功能验证实验和对小鼠疼痛行为的分析表明，确实发现了有意义的蛋白质变化。这些结果说明了DIA-MS的应用如何为实现期待已久的体感系统病理分子解剖的标准化开辟了新的途径。因此，我们的发现提供了一个有价值的框架，以定性地扩展我们对慢性疼痛和躯体感觉的理解。
Chronic pain is a complex disease with limited treatment options. Several profiling efforts have been employed with the aim to dissect its molecular underpinnings. However, generated results are often inconsistent and nonoverlapping, which is largely because of inherent technical constraints. Emerging data-independent acquisition (DIA)-mass spectrometry (MS) has the potential to provide unbiased, reproducible and quantitative proteome maps - a prerequisite for standardization among experiments. Here, we designed a DIA-based proteomics workflow to profile changes in the abundance of dorsal root ganglia (DRG) proteins in two mouse models of chronic pain, inflammatory and neuropathic. We generated a DRG-specific spectral library containing 3067 DRG proteins, which enables their standardized quantification by means of DIA-MS in any laboratory. Using this resource, we profiled 2526 DRG proteins in each biological replicate of both chronic pain models and respective controls with unprecedented reproducibility. We detected numerous differentially regulated proteins, the majority of which exhibited pain model-specificity. Our approach recapitulates known biology and discovers dozens of proteins that have not been characterized in the somatosensory system before. Functional validation experiments and analysis of mouse pain behaviors demonstrate that indeed meaningful protein alterations were discovered. These results illustrate how the application of DIA-MS can open new avenues to achieve the long-awaited standardization in the molecular dissection of pathologies of the somatosensory system. Therefore, our findings provide a valuable framework to qualitatively extend our understanding of chronic pain and somatosensation.