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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

The process of individual SRV9 transport in a Vero cell.Vero cells were infected with SRV9 (red), and imaged 10 min after the infection. The nucleus region was circled by blue dashed line. The white arrow points a single virus inside the cell. The trajectory of a Cy5-labeled virus inside a cell is shown by yellow line. The movement of the virus was recorded by real-time confocal microscopy. The time interval of serial images was 10 s. Scale bar: 20 μm.
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f5: The process of individual SRV9 transport in a Vero cell.Vero cells were infected with SRV9 (red), and imaged 10 min after the infection. The nucleus region was circled by blue dashed line. The white arrow points a single virus inside the cell. The trajectory of a Cy5-labeled virus inside a cell is shown by yellow line. The movement of the virus was recorded by real-time confocal microscopy. The time interval of serial images was 10 s. Scale bar: 20 μm.

Mentions: Next, to further reveal the infection process, we utilized the real-time tracking to monitor the individual virus transport in live cells. As shown in Fig. 5, a single virus was imaged in the cell as pointed by white arrow, and the cell nucleus was circled by the blue dashed line. The typical trajectory of a virus starting in the cell periphery, moving to the perinuclear region and finally locating there is shown in Fig. 5. The rapid movement process is recorded in Movie S5 in Supplementary Information. In short, the single-virus tracking is an excellent tool to follow the fate of individual virus particle in live cells47. By means of this technique, for the first time, we directly revealed characteristics of the internalization and transport behavior of single SRV9 particle in living cells, which is significant for understanding the initial stage of viral infection process.


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)

The process of individual SRV9 transport in a Vero cell.Vero cells were infected with SRV9 (red), and imaged 10 min after the infection. The nucleus region was circled by blue dashed line. The white arrow points a single virus inside the cell. The trajectory of a Cy5-labeled virus inside a cell is shown by yellow line. The movement of the virus was recorded by real-time confocal microscopy. The time interval of serial images was 10 s. Scale bar: 20 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: The process of individual SRV9 transport in a Vero cell.Vero cells were infected with SRV9 (red), and imaged 10 min after the infection. The nucleus region was circled by blue dashed line. The white arrow points a single virus inside the cell. The trajectory of a Cy5-labeled virus inside a cell is shown by yellow line. The movement of the virus was recorded by real-time confocal microscopy. The time interval of serial images was 10 s. Scale bar: 20 μm.
Mentions: Next, to further reveal the infection process, we utilized the real-time tracking to monitor the individual virus transport in live cells. As shown in Fig. 5, a single virus was imaged in the cell as pointed by white arrow, and the cell nucleus was circled by the blue dashed line. The typical trajectory of a virus starting in the cell periphery, moving to the perinuclear region and finally locating there is shown in Fig. 5. The rapid movement process is recorded in Movie S5 in Supplementary Information. In short, the single-virus tracking is an excellent tool to follow the fate of individual virus particle in live cells47. By means of this technique, for the first time, we directly revealed characteristics of the internalization and transport behavior of single SRV9 particle in living cells, which is significant for understanding the initial stage of viral infection process.

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