Limits...
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 correlation of the viruses with filopodia.(A) The confocal images of Vero cells (green) that unexposed or exposed to SRV9. (B) The number of filopodia per cell with or without SRV9 exposure was quantified. Error bars represent standard error of the mean (SEM). (C) The average length of filopodia with or without SRV9 exposure was quantified. Error bars represent SEM. In D and E, SRV9 (red) were added to Vero cells (green) and immediately monitored over time by confocal fluorescence microscopy. (D) Arrowheads mark the movement of single virus particles via retrograde transport from the filopodium periphery toward the cell body. (E) An example of single virus particles reaching the cell body through filopodial retraction where the filopodial tip is marked by arrowheads. Scale bars: 10 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4493577&req=5

f2: The correlation of the viruses with filopodia.(A) The confocal images of Vero cells (green) that unexposed or exposed to SRV9. (B) The number of filopodia per cell with or without SRV9 exposure was quantified. Error bars represent standard error of the mean (SEM). (C) The average length of filopodia with or without SRV9 exposure was quantified. Error bars represent SEM. In D and E, SRV9 (red) were added to Vero cells (green) and immediately monitored over time by confocal fluorescence microscopy. (D) Arrowheads mark the movement of single virus particles via retrograde transport from the filopodium periphery toward the cell body. (E) An example of single virus particles reaching the cell body through filopodial retraction where the filopodial tip is marked by arrowheads. Scale bars: 10 μm.

Mentions: To observe the association of SRV9 with the filopodia, we implemented the following contrasting experiment. Visualization of the Vero cells in unexposed SRV9 revealed few filopodia (Fig. 2A; mock). Notably, after cells exposured to SRV9 for short periods of time, a strong induction of filopodia was observed (Fig. 2A; +SRV9). Simultaneously, we quantitatively analyzed the effect of the SRV9 exposure on the number of filopodia per cell and the average length of filopodia in more than 100 cells. (Fig. 2B,C). The results indicated SRV9 addition has activated more filopodia production and elongation, which may induce increased virus uptake from the surrounding environment.


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 correlation of the viruses with filopodia.(A) The confocal images of Vero cells (green) that unexposed or exposed to SRV9. (B) The number of filopodia per cell with or without SRV9 exposure was quantified. Error bars represent standard error of the mean (SEM). (C) The average length of filopodia with or without SRV9 exposure was quantified. Error bars represent SEM. In D and E, SRV9 (red) were added to Vero cells (green) and immediately monitored over time by confocal fluorescence microscopy. (D) Arrowheads mark the movement of single virus particles via retrograde transport from the filopodium periphery toward the cell body. (E) An example of single virus particles reaching the cell body through filopodial retraction where the filopodial tip is marked by arrowheads. Scale bars: 10 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: The correlation of the viruses with filopodia.(A) The confocal images of Vero cells (green) that unexposed or exposed to SRV9. (B) The number of filopodia per cell with or without SRV9 exposure was quantified. Error bars represent standard error of the mean (SEM). (C) The average length of filopodia with or without SRV9 exposure was quantified. Error bars represent SEM. In D and E, SRV9 (red) were added to Vero cells (green) and immediately monitored over time by confocal fluorescence microscopy. (D) Arrowheads mark the movement of single virus particles via retrograde transport from the filopodium periphery toward the cell body. (E) An example of single virus particles reaching the cell body through filopodial retraction where the filopodial tip is marked by arrowheads. Scale bars: 10 μm.
Mentions: To observe the association of SRV9 with the filopodia, we implemented the following contrasting experiment. Visualization of the Vero cells in unexposed SRV9 revealed few filopodia (Fig. 2A; mock). Notably, after cells exposured to SRV9 for short periods of time, a strong induction of filopodia was observed (Fig. 2A; +SRV9). Simultaneously, we quantitatively analyzed the effect of the SRV9 exposure on the number of filopodia per cell and the average length of filopodia in more than 100 cells. (Fig. 2B,C). The results indicated SRV9 addition has activated more filopodia production and elongation, which may induce increased virus uptake from the surrounding environment.

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