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Real-time Imaging of Rabies Virus Entry into Living Vero cells.

Xu H, Hao X, Wang S, Wang Z, Cai M, Jiang J, Qin Q, Zhang M, Wang H - Sci Rep (2015)

Bottom Line: Firstly, it was found that the actin-enriched filopodia is in favor of virus reaching to the cell body.Then, our real-time imaging results unambiguously uncover the characteristics of viral internalization and cellular transport dynamics.Significantly, the results provide profound insight into development of novel and effective antiviral targets.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, P.R. China.

ABSTRACT
Understanding the mechanism of rabies virus (RABV) infection is vital for prevention and therapy of virulent rabies. However, the infection mechanism remains largely uncharacterized due to the limited methods and viral models. Herein, we utilized a powerful single-virus tracking technique to dynamically and globally visualize the infection process of the live attenuated rabies vaccine strain-SRV9 in living Vero cells. Firstly, it was found that the actin-enriched filopodia is in favor of virus reaching to the cell body. Furthermore, by carrying out drug perturbation experiments, we confirmed that RABV internalization into Vero cells proceeds via classical dynamin-dependent clathrin-mediated endocytosis with requirement for intact actin, but caveolae-dependent endocytosis is not involved. Then, our real-time imaging results unambiguously uncover the characteristics of viral internalization and cellular transport dynamics. In addition, our results directly and quantitatively reveal that the intracellular motility of internalized RABV particles is largely microtubule-dependent. Collectively, our work is crucial for understanding the initial steps of RABV infection, and elucidating the mechanisms of post-infection. Significantly, the results provide profound insight into development of novel and effective antiviral targets.

No MeSH data available.


Related in: MedlinePlus

Colocalization of SRV9 particles with microtubules.Immunofluorescence was performed at cells infected by Cy5-labele SRV9 (red) after 30 min using anti-tublin antibody (green). The white square region is magnified and shown in the inset. The white arrows represent some examples of colocalization of Cy5-labeled SRV9 with FITC-tagged microtubules. Scale bar: 10 μm.
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f7: Colocalization of SRV9 particles with microtubules.Immunofluorescence was performed at cells infected by Cy5-labele SRV9 (red) after 30 min using anti-tublin antibody (green). The white square region is magnified and shown in the inset. The white arrows represent some examples of colocalization of Cy5-labeled SRV9 with FITC-tagged microtubules. Scale bar: 10 μm.

Mentions: Next, we performed the colocalization experiment to analyze the involvement of microtubules in the intracellular motility of SRV9 (Fig. 7). The microtubules were recognized by antibody against tubulin. In the magnified image (inset), these yellow signals as indicated by white arrows represented the overlap of the microtubules (green) and Cy5-labeled SRV9 (red), indicating that some SRV9 colocalized with the microtubules. Our statistical result indicated that about 77.6% of SRV9 virions are colocalized with microtubules at 30 min for virus infection. In addition, SRV9 particles were observed to move along microtubules during infection (see Movie S8 in the Supplementary Information). The results suggested that microtubules may be crucial for virus transport during infection.


Real-time Imaging of Rabies Virus Entry into Living Vero cells.

Xu H, Hao X, Wang S, Wang Z, Cai M, Jiang J, Qin Q, Zhang M, Wang H - Sci Rep (2015)

Colocalization of SRV9 particles with microtubules.Immunofluorescence was performed at cells infected by Cy5-labele SRV9 (red) after 30 min using anti-tublin antibody (green). The white square region is magnified and shown in the inset. The white arrows represent some examples of colocalization of Cy5-labeled SRV9 with FITC-tagged microtubules. Scale bar: 10 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493577&req=5

f7: Colocalization of SRV9 particles with microtubules.Immunofluorescence was performed at cells infected by Cy5-labele SRV9 (red) after 30 min using anti-tublin antibody (green). The white square region is magnified and shown in the inset. The white arrows represent some examples of colocalization of Cy5-labeled SRV9 with FITC-tagged microtubules. Scale bar: 10 μm.
Mentions: Next, we performed the colocalization experiment to analyze the involvement of microtubules in the intracellular motility of SRV9 (Fig. 7). The microtubules were recognized by antibody against tubulin. In the magnified image (inset), these yellow signals as indicated by white arrows represented the overlap of the microtubules (green) and Cy5-labeled SRV9 (red), indicating that some SRV9 colocalized with the microtubules. Our statistical result indicated that about 77.6% of SRV9 virions are colocalized with microtubules at 30 min for virus infection. In addition, SRV9 particles were observed to move along microtubules during infection (see Movie S8 in the Supplementary Information). The results suggested that microtubules may be crucial for virus transport during infection.

Bottom Line: Firstly, it was found that the actin-enriched filopodia is in favor of virus reaching to the cell body.Then, our real-time imaging results unambiguously uncover the characteristics of viral internalization and cellular transport dynamics.Significantly, the results provide profound insight into development of novel and effective antiviral targets.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, P.R. China.

ABSTRACT
Understanding the mechanism of rabies virus (RABV) infection is vital for prevention and therapy of virulent rabies. However, the infection mechanism remains largely uncharacterized due to the limited methods and viral models. Herein, we utilized a powerful single-virus tracking technique to dynamically and globally visualize the infection process of the live attenuated rabies vaccine strain-SRV9 in living Vero cells. Firstly, it was found that the actin-enriched filopodia is in favor of virus reaching to the cell body. Furthermore, by carrying out drug perturbation experiments, we confirmed that RABV internalization into Vero cells proceeds via classical dynamin-dependent clathrin-mediated endocytosis with requirement for intact actin, but caveolae-dependent endocytosis is not involved. Then, our real-time imaging results unambiguously uncover the characteristics of viral internalization and cellular transport dynamics. In addition, our results directly and quantitatively reveal that the intracellular motility of internalized RABV particles is largely microtubule-dependent. Collectively, our work is crucial for understanding the initial steps of RABV infection, and elucidating the mechanisms of post-infection. Significantly, the results provide profound insight into development of novel and effective antiviral targets.

No MeSH data available.


Related in: MedlinePlus