Comprehensive Proteome Analysis of Hippocampus, Brainstem, and Spinal Cord from Paralytic and Furious Dogs Naturally Infected with Rabies

Abstract

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.