Quantitative Proteome Profiling of Street Rabies Virus-Infected Mouse Hippocampal Synaptosomes

Abstract

It is well established now that neuronal dysfunction rather than structural damage may be responsible for the development of rabies. In order to explore the underlying mechanisms in rabies virus (RABV) and synaptic dysfunctions, a quantitative proteome profiling was carried out on synaptosome samples from mice hippocampus. Synaptosome samples from mice hippocampus were isolated and confirmed by Western blot and transmission electron microscopy. Synaptosome protein content changes were quantitatively detected by Nano-LC-MS/MS. Protein functions were classified by the Gene Ontology (GO) and KEGG pathway. PSICQUIC was used to create a network. MCODE algorithm was applied to obtain subnetworks. Of these protein changes, 45 were upregulated and 14 were downregulated following RABV infection relative to non-infected (mock) synaptosomes. 28 proteins were unique to mock treatment and 12 were unique to RABV treatment. Proteins related to metabolism and synaptic vesicle showed the most changes in expression levels. Furthermore, protein-protein interaction (PPI) networks revealed that several key biological processes related to synaptic functions potentially were modulated by RABV, including energy metabolism, cytoskeleton organization, and synaptic transmission. These data will be useful for better understanding of neuronal dysfunction of rabies and provide the foundation for future research.

Rabies Virus Exploits Cytoskeleton Network to Cause Early Disease Progression and Cellular Dysfunction

“…The vimentin belongs to the intermediate filaments of the cytoskeleton, and preserves the cellular morphology by maintaining cytoskeleton shape, while tropomyosin also adjusts muscular contractions with the help of actin filaments ( 4 , 14 ). Few studies of differential gene expression describe the abnormal neuronal structures due to the aberrations in the cytoplasmic proteins, and their corresponding pathways under RABV infections ( 22 – 24 ). Of these proteins, actin-related protein 2/3 complex subunit 3 and actin-related protein 2 showed higher gene expression, and the first one is meant for the actin polymerization, while the latter one maintains cellular polarity through the organization of the cytoskeleton in synaptosomes ( 24 ).…”

“…Few studies of differential gene expression describe the abnormal neuronal structures due to the aberrations in the cytoplasmic proteins, and their corresponding pathways under RABV infections ( 22 – 24 ). Of these proteins, actin-related protein 2/3 complex subunit 3 and actin-related protein 2 showed higher gene expression, and the first one is meant for the actin polymerization, while the latter one maintains cellular polarity through the organization of the cytoskeleton in synaptosomes ( 24 ). However, neurofilament light polypeptide shows lower gene expression because it controls the axonal transport and aligns the assembly of axonal bundles ( 24 ).…”

“…Of these proteins, actin-related protein 2/3 complex subunit 3 and actin-related protein 2 showed higher gene expression, and the first one is meant for the actin polymerization, while the latter one maintains cellular polarity through the organization of the cytoskeleton in synaptosomes ( 24 ). However, neurofilament light polypeptide shows lower gene expression because it controls the axonal transport and aligns the assembly of axonal bundles ( 24 ). These are the salient proteins, and yet there are other actin-associated proteins (alpha-actinin-1 that anchors actin to various cellular structures, dynein light chain (DLC), capping proteins, and drebrin that generate stabilized actin filaments concentrated in the dendritic spines) and microtubule-associated proteins (microtubule-associated proteins-6) which are abundantly concentrated in the synaptosomes ( 5 , 15 , 24 ).…”

What Kaplan-Meier survival curves don't tell us about CNS disease

“…In fact, many viruses, including those with RNA genomes, can be maintained long after infectious virus falls below detection limits, and in the absence of overt signs of CNS disease [21][22][23] . Such long-lasting infections, while not lethal to the host, could nevertheless impair neuronal function and host metabolism 24,25 . In some instances, viral persistence correlates with obvious neurological disease; for example, after infection with mouse hepatitis virus (MHV), mice develop a progressive demyelinating disease that shares many attributes with multiple sclerosis 26 .…”

Establishment of Human-Induced Pluripotent Stem Cell-Derived Neurons—A Promising In Vitro Model for a Molecular Study of Rabies Virus and Host Interaction

“…Each approach has strengths and limitations in applying biological data to answer specific research questions. The proteomics-based analysis is a powerful platform to predict and identify cellular mechanisms during RABV infection in permissive cell lines and infected human and animal brain tissues [ 23 , 24 , 25 , 26 ]. Despite many relevant protein profiles reported in RABV-infected BHK21 and neuroblastoma cell lines, the relevance of the identified proteins in these non-neuronal cell lines is obscure [ 27 , 28 ].…”