Inter-Department Program of Biomedical Sciences, Chulalongkorn University, Bangkok, Thailand.
J Proteome Res. 2011 Nov 4;10(11):4911-24. doi: 10.1021/pr200276u. Epub 2011 Oct 18.
Paralytic and furious forms are unique clinical entities of rabies in humans and dogs. However, molecular mechanisms underlying these disorders remained unclear. We investigated changes in proteomes of the hippocampus, brainstem and spinal cord of paralytic and furious dogs naturally infected with rabies compared to noninfected controls. Proteins were extracted from these tissues and analyzed by two-dimensional gel electrophoresis (2-DE) (n = 6 gels/region in each group, a total of 54 gels were analyzed). From >1000 protein spots visualized in each gel, spot matching, quantitative intensity analysis, and ANOVA with Tukey's posthoc multiple comparisons revealed 32, 49, and 67 protein spots that were differentially expressed among the three clinical groups in the hippocampus, brainstem and spinal cord, respectively. These proteins were then identified by quadrupole time-of-flight mass spectrometry and tandem mass spectrometry (Q-TOF MS and MS/MS), including antioxidants, apoptosis-related proteins, cytoskeletal proteins, heat shock proteins/chaperones, immune regulatory proteins, metabolic enzymes, neuron-specific proteins, transcription/translation regulators, ubiquitination/proteasome-related proteins, vesicular transport proteins, and hypothetical proteins. Among these, 13, 17, and 41 proteins in the hippocampus, brainstem and spinal cord, respectively, significantly differed between paralytic and furious forms and thus may potentially be biomarkers to differentiate these two distinct forms of rabies. In summary, we report herein for the first time a large data set of changes in proteomes of the hippocampus, brainstem and spinal cord in dogs naturally infected with rabies. These data will be useful for better understanding of molecular mechanisms of rabies and for differentiation of its paralytic and furious forms.
麻痹型和狂躁型是人类和犬类狂犬病的独特临床实体。然而，这些疾病的分子机制尚不清楚。我们研究了自然感染狂犬病的麻痹型和狂躁型犬的海马体、脑干和脊髓的蛋白质组的变化，并与未感染的对照组进行了比较。从这些组织中提取蛋白质，并通过二维凝胶电泳（2-DE）进行分析（每组 6 个凝胶/区域，共分析了 54 个凝胶）。在每个凝胶中可视化的 >1000 个蛋白斑点中，斑点匹配、定量强度分析和方差分析与 Tukey 的事后多重比较显示，在海马体、脑干和脊髓中，这三个临床组之间有 32、49 和 67 个蛋白斑点存在差异表达。然后通过四极飞行时间质谱和串联质谱（Q-TOF MS 和 MS/MS）对这些蛋白进行鉴定，包括抗氧化剂、凋亡相关蛋白、细胞骨架蛋白、热休克蛋白/伴侣蛋白、免疫调节蛋白、代谢酶、神经元特异性蛋白、转录/翻译调节因子、泛素化/蛋白酶体相关蛋白、囊泡运输蛋白和假设蛋白。其中，海马体、脑干和脊髓中的 13、17 和 41 种蛋白在麻痹型和狂躁型之间存在显著差异，因此可能是区分这两种不同形式狂犬病的潜在生物标志物。总之，我们首次报道了自然感染狂犬病的犬海马体、脑干和脊髓蛋白质组变化的大型数据集。这些数据将有助于更好地理解狂犬病的分子机制，并有助于区分其麻痹型和狂躁型。
