Differential expression of proteins in the central nervous system of early and late stage rabies infected dogs

Rabies is a zoonotic disease. Dogs are an important reservoir host of rabies in Thailand, Asia, Africa and Europe. The clinical manifestations are the same in humans and dogs, and are categorized into 2 forms; furious and paralytic. Although there have been many rabies studies over the years, the pathogenesis of rabies is still unclear. In this study, proteomic analysis was used to investigate changes in host responses in central nervous system (CNS) tissues at early and late stages of disease compared to non-infected dog controls. They were divided in 5 groups; early furious (FE), late furious (FL), early paralytic (DE), late paralytic (DL) and control (N), respectively. Hippocampus, parietal lobes, brainstem and spinal cord tissues from these 5 groups were collected and stored until examination. Proteins were extracted from these tissues and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in combination with liquid chromatography mass spectrometry (LC-MS/MS). The proteins were statistically selected by suitable bioinformatics tools. There were 26, 7, 35 and 18 proteins expressed only in each hippocampus, parietal lobes, brainstem and spinal cord, respectively. They played role in cytoskeleton proteins, enzymes, proteins associated with cell death, inflammation and immunity. This is the first report of changes in proteome data set from rabies-infected CNS tissues in both forms, and in early and late stages of infection. This data will be useful for not only better understanding of the molecular mechanism in many regions of CNS tissues, but also diagnostic and therapeutic of rabies.

狂犬病是一种人畜共患病。犬类是泰国、亚洲、非洲及欧洲地区狂犬病的重要储存宿主。人与犬的临床症状表现一致，可分为两种类型：狂暴型（furious）与麻痹型（paralytic）。尽管多年来已有诸多狂犬病相关研究，但狂犬病的发病机制仍未明确。本研究采用蛋白质组学分析方法，探究感染狂犬病犬在疾病早期与晚期阶段，其中枢神经系统（central nervous system, CNS）组织内宿主应答的变化，并以未感染犬作为对照。实验对象分为5组，分别为早期狂暴型（FE）、晚期狂暴型（FL）、早期麻痹型（DE）、晚期麻痹型（DL）以及对照组（N）。采集上述5组犬的海马体、顶叶、脑干及脊髓组织并保存，待后续检测。从上述组织中提取蛋白质，采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE）结合液相色谱-串联质谱（liquid chromatography mass spectrometry, LC-MS/MS）进行分析。通过合适的生物信息学工具对蛋白质进行统计学筛选。仅在海马体、顶叶、脑干及脊髓中特异性表达的蛋白质分别有26种、7种、35种及18种，这些蛋白质涉及细胞骨架蛋白、酶类以及与细胞死亡、炎症和免疫相关的功能。本研究首次报道了涵盖两种临床类型、不同感染阶段的狂犬病感染中枢神经系统组织的蛋白质组变化数据集。该数据集不仅有助于深入解析中枢神经系统不同区域的分子致病机制，还可为狂犬病的诊断与治疗提供参考依据。