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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

(A,B) The histogram of motility speed distribution of SRV9 in the untreated and nocodazole-treated Vero cells, respectively. (C) Comparison of SRV9 motility with the speed >0.05 μm s−1 between the untreated and nocodazole-treated Vero cells.
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f8: (A,B) The histogram of motility speed distribution of SRV9 in the untreated and nocodazole-treated Vero cells, respectively. (C) Comparison of SRV9 motility with the speed >0.05 μm s−1 between the untreated and nocodazole-treated Vero cells.

Mentions: Because of complexity and heterogeneity of the intracellular environment, the viruses can exploit more than one transport system simultaneously to efficiently infect their hosts4849. Therefore, it is necessary to characterize and quantitatively analyze the viral motility by tracking a large number of virus particles in cells. The statistical analyses were implemented on tens of thousands of trajectories in many independent experiments. The Fig. 8A,B presented statistics histogram of speed in untreated cell and nocodazole-treated cell, respectively. The statistical analysis from the two group cells is shown in Fig. 8C, displaying that the percentage of trajectories with peak speeds >0.05 μm s−1 is 24.7% in untreated cells, and 5.9% in nocodazole-treated cells. The 4.2-fold reduction in the speed of SRV9 motility occurred in microtubule-depolymerized cells. Therefore, we speculated that SRV9 motility was closely related to the microtubules in living cells. Recently, the microtubule-mediated behavior of virus motility has been proven as a complex event50. Future work will be required to determine whether this microtubule-associated motility way ultimately contributes to productive infection, and if so what underlying mechanism is involved.


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)

(A,B) The histogram of motility speed distribution of SRV9 in the untreated and nocodazole-treated Vero cells, respectively. (C) Comparison of SRV9 motility with the speed >0.05 μm s−1 between the untreated and nocodazole-treated Vero cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f8: (A,B) The histogram of motility speed distribution of SRV9 in the untreated and nocodazole-treated Vero cells, respectively. (C) Comparison of SRV9 motility with the speed >0.05 μm s−1 between the untreated and nocodazole-treated Vero cells.
Mentions: Because of complexity and heterogeneity of the intracellular environment, the viruses can exploit more than one transport system simultaneously to efficiently infect their hosts4849. Therefore, it is necessary to characterize and quantitatively analyze the viral motility by tracking a large number of virus particles in cells. The statistical analyses were implemented on tens of thousands of trajectories in many independent experiments. The Fig. 8A,B presented statistics histogram of speed in untreated cell and nocodazole-treated cell, respectively. The statistical analysis from the two group cells is shown in Fig. 8C, displaying that the percentage of trajectories with peak speeds >0.05 μm s−1 is 24.7% in untreated cells, and 5.9% in nocodazole-treated cells. The 4.2-fold reduction in the speed of SRV9 motility occurred in microtubule-depolymerized cells. Therefore, we speculated that SRV9 motility was closely related to the microtubules in living cells. Recently, the microtubule-mediated behavior of virus motility has been proven as a complex event50. Future work will be required to determine whether this microtubule-associated motility way ultimately contributes to productive infection, and if so what underlying mechanism is involved.

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