Pathogenic and attenuated rabies viruses induces differential host protein expression in the central

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Pathogenic and attenuated rabies viruses induces
differential host protein expression in the
central nervous system Implication of neuronal
dysfunction

• Zhen F. Fu
• Department of Pathology
• University of Georgia

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Rabies Pathogenesis
Robert Hurt-USC
Patients die of circulatory insufficiency,
cardiac arrest and respiratory failure.
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Despite extensive research in the past 100 years,
we still know very little about the pathogenic
mechanism by which rabies virus infection of
neurons causes rabies.There are scarce
neuropathology with mild inflammation and little
neuronal loss, which cannot explain the lethality
of the disease.It has been hypothesized that
rabies results from neuronal dysfunction rather
than structural damage. However, it is not known
how RV infection leads to neuronal
dysfunction.To better understand rabies
pathogenesis, we initiate a project to determine
how the host responds to rabies virus infections
using one street and one fixed virus. This is
accomplished by using proteomics technologies.
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Two viruses were used in this studySHBRV Wt
virus, normally circulating in silver-haired bats
and responsible for most of the human rabies in
the US.CVS-B2C Lab-adapted attenuated virus
derived from CVS-24 by passaging in BHK cells.
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Survival curve
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Detection of Differential Protein Levels in the
Proteome
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mutant/Infected
Wild type
In-gel digestion with protease Identify the
protein by Mass Spectrometry
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Identification of gel-separated proteins by mass
spectrometry
Gygi et al.
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The situation to avoid
Software is essential !
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Proteins differentially expressed in response to
SHBRV infection
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Proteins differentially expressed in response to
B2C infection in mice
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Western blotting of proteins involved in ion
homeostasis and synaptic physiology
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Accumulation of vesicles
No Docking
No release of Neurotransmitters
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SHBRV B2C Control
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Conclusions
Proteomics data indicate that wt RV infection
resulted in up-regulation of proteins involved in
ion homeostasis and down-regulation of synaptic
proteins. The altered protein expression as
detected by 2D-gel analysis is confirmed by
Western blotting in animals infected either ic or
im as well as in primary neuron. Up-regulation
of Na/K-ATPase leads to decrease in Na
concentrations in infected cells. Likewise,
down-regulation of Ca-ATPase resulted in decrease
of Ca concentration in infected cells. Changes
in Na/Ca concentration affects membrane potential
and thus leading to alteration of neuronal
transmission. Synaptic proteins such as
syntaxin, a-SANP, and TRIM9 play important roles
in synaptic-vesicle fusion and docking of
synaptic vesicles. Down-regulation of these
proteins prevented the docking and fusion of
synaptic vesicles with presynaptic membrane, thus
resulting in accumulation of synaptic vesicles in
the presynapses. Thus our data may provide
structural and metabolic basis by which RV
infection causes neuronal dysfunction.
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Acknowledgements Vikas Dhingra Xia-qin
g Li Luciana Sarmento UGA
Proteomics Facility Tracy Andachtc

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Thank you!!