Frontiers in Imaging

A special issue of Viruses (ISSN 1999-4915). This special issue belongs to the section "Animal Viruses".

Deadline for manuscript submissions: closed (31 August 2012) | Viewed by 39567

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Department of Cell & Molecular Biology, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA

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Department of Microbiology and Immunology, Stritch School of Medicine, Loyola University, Chicago, IL, USA

Published Papers (5 papers)

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Research

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2095 KiB  
Article
High Throughput Method to Quantify Anterior-Posterior Polarity of T-Cells and Epithelial Cells
by Charletha V. Irvin-Wilson, Justin Y. Newberg, Kathleen Kong, Ronald T. Javier and Susan J. Marriott
Viruses 2011, 3(12), 2396-2411; https://doi.org/10.3390/v3122396 - 28 Nov 2011
Viewed by 6591
Abstract
The virologic synapse (VS), which is formed between a virus-infected and uninfected cell, plays a central role in the transmission of certain viruses, such as HIV and HTLV-1. During VS formation, HTLV-1-infected T-cells polarize cellular and viral proteins toward the uninfected T-cell. This [...] Read more.
The virologic synapse (VS), which is formed between a virus-infected and uninfected cell, plays a central role in the transmission of certain viruses, such as HIV and HTLV-1. During VS formation, HTLV-1-infected T-cells polarize cellular and viral proteins toward the uninfected T-cell. This polarization resembles anterior-posterior cell polarity induced by immunological synapse (IS) formation, which is more extensively characterized than VS formation and occurs when a T-cell interacts with an antigen-presenting cell. One measure of cell polarity induced by both IS or VS formation is the repositioning of the microtubule organizing center (MTOC) relative to the contact point with the interacting cell. Here we describe an automated, high throughput system to score repositioning of the MTOC and thereby cell polarity establishment. The method rapidly and accurately calculates the angle between the MTOC and the IS for thousands of cells. We also show that the system can be adapted to score anterior-posterior polarity establishment of epithelial cells. This general approach represents a significant advancement over manual cell polarity scoring, which is subject to experimenter bias and requires more time and effort to evaluate large numbers of cells. Full article
(This article belongs to the Special Issue Frontiers in Imaging)
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Review

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488 KiB  
Review
Application of Live-Cell RNA Imaging Techniques to the Study of Retroviral RNA Trafficking
by Darrin V. Bann and Leslie J. Parent
Viruses 2012, 4(6), 963-979; https://doi.org/10.3390/v4060963 - 08 Jun 2012
Cited by 21 | Viewed by 10460
Abstract
Retroviruses produce full-length RNA that serves both as a genomic RNA (gRNA), which is encapsidated into virus particles, and as an mRNA, which directs the synthesis of viral structural proteins. However, we are only beginning to understand the cellular and viral factors that [...] Read more.
Retroviruses produce full-length RNA that serves both as a genomic RNA (gRNA), which is encapsidated into virus particles, and as an mRNA, which directs the synthesis of viral structural proteins. However, we are only beginning to understand the cellular and viral factors that influence trafficking of retroviral RNA and the selection of the RNA for encapsidation or translation. Live cell imaging studies of retroviral RNA trafficking have provided important insight into many aspects of the retrovirus life cycle including transcription dynamics, nuclear export of viral RNA, translational regulation, membrane targeting, and condensation of the gRNA during virion assembly. Here, we review cutting-edge techniques to visualize single RNA molecules in live cells and discuss the application of these systems to studying retroviral RNA trafficking. Full article
(This article belongs to the Special Issue Frontiers in Imaging)
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2056 KiB  
Review
Quantitative Live-Cell Imaging of Human Immunodeficiency Virus (HIV-1) Assembly
by Viola Baumgärtel, Barbara Müller and Don C. Lamb
Viruses 2012, 4(5), 777-799; https://doi.org/10.3390/v4050777 - 04 May 2012
Cited by 21 | Viewed by 9402
Abstract
Advances in fluorescence methodologies make it possible to investigate biological systems in unprecedented detail. Over the last few years, quantitative live-cell imaging has increasingly been used to study the dynamic interactions of viruses with cells and is expected to become even more indispensable [...] Read more.
Advances in fluorescence methodologies make it possible to investigate biological systems in unprecedented detail. Over the last few years, quantitative live-cell imaging has increasingly been used to study the dynamic interactions of viruses with cells and is expected to become even more indispensable in the future. Here, we describe different fluorescence labeling strategies that have been used to label HIV-1 for live cell imaging and the fluorescence based methods used to visualize individual aspects of virus-cell interactions. This review presents an overview of experimental methods and recent experiments that have employed quantitative microscopy in order to elucidate the dynamics of late stages in the HIV-1 replication cycle. This includes cytosolic interactions of the main structural protein, Gag, with itself and the viral RNA genome, the recruitment of Gag and RNA to the plasma membrane, virion assembly at the membrane and the recruitment of cellular proteins involved in HIV-1 release to the nascent budding site. Full article
(This article belongs to the Special Issue Frontiers in Imaging)
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2513 KiB  
Review
The Use of Fluorescence Microscopy to Study the Association Between Herpesviruses and Intrinsic Resistance Factors
by Roger D. Everett
Viruses 2011, 3(12), 2412-2424; https://doi.org/10.3390/v3122412 - 07 Dec 2011
Cited by 7 | Viewed by 6761
Abstract
Intrinsic antiviral resistance is a branch of antiviral defence that involves constitutively expressed cellular proteins that act within individual infected cells. In recent years it has been discovered that components of cellular nuclear structures known as ND10 or PML nuclear bodies contribute to [...] Read more.
Intrinsic antiviral resistance is a branch of antiviral defence that involves constitutively expressed cellular proteins that act within individual infected cells. In recent years it has been discovered that components of cellular nuclear structures known as ND10 or PML nuclear bodies contribute to intrinsic resistance against a variety of viruses, notably of the herpesvirus family. Several ND10 components are rapidly recruited to sites that are closely associated with herpes simplex virus type 1 (HSV-1) genomes during the earliest stages of infection, and this property correlates with the efficiency of ND10 mediated restriction of HSV-1 replication. Similar but distinct recruitment of certain DNA damage response proteins also occurs during infection. These recruitment events are inhibited in a normal wild type HSV-1 infection by the viral regulatory protein ICP0. HSV‑1 mutants that do not express ICP0 are highly susceptible to repression through intrinsic resistance factors, but they replicate more efficiently in cells depleted of certain ND10 proteins or in which ND10 component recruitment is inefficient. This article presents the background to this recruitment phenomenon and summaries how it is conveniently studied by fluorescence microscopy. Full article
(This article belongs to the Special Issue Frontiers in Imaging)
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2030 KiB  
Review
Hepatitis C Virus Assembly Imaging
by Costin-Ioan Popescu, Yves Rouillé and Jean Dubuisson
Viruses 2011, 3(11), 2238-2254; https://doi.org/10.3390/v3112238 - 15 Nov 2011
Cited by 17 | Viewed by 5957
Abstract
Hepatitis C Virus (HCV) assembly process is the least understood step in the virus life cycle. The functional data revealed by forward and reverse genetics indicated that both structural and non-structural proteins are involved in the assembly process. Using confocal and electron microscopy [...] Read more.
Hepatitis C Virus (HCV) assembly process is the least understood step in the virus life cycle. The functional data revealed by forward and reverse genetics indicated that both structural and non-structural proteins are involved in the assembly process. Using confocal and electron microscopy different groups determined the subcellular localization of different viral proteins and they identified the lipid droplets (LDs) as the potential viral assembly site. Here, we aim to review the mechanisms that govern the viral proteins recruitment to LDs and discuss the current model of HCV assembly process. Based on previous examples, this review will also discuss advanced imaging techniques as potential means to extend our present knowledge of HCV assembly process. Full article
(This article belongs to the Special Issue Frontiers in Imaging)
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