University of Southern California, Los Angeles

6 ARTICLES PUBLISHED IN JoVE

Biology

Studying Age-dependent Genomic Instability using the S. cerevisiae Chronological Lifespan Model

Min Wei¹, Federica Madia¹, Valter D. Longo¹

¹Andrus Gerontology Center, Department of Biological Sciences, Department of Molecular and Computational Biology, University of Southern California, Los Angeles

Here we describe a set of DNA mutation assays that can be combined with the yeast chronological life span model to study the genes/pathways that regulate or contribute to genomic DNA instability during aging.

Immunology and Infection

Measurement of γHV68 Infection in Mice

Sara Dolatshahi Pirooz¹, Joo-Hyung Lee¹, Zhen Zhao¹, Duojiao Ni¹, Soohwan Oh¹, Chengyu Liang¹

¹Department of Molecular Microbiology and Immunology, University of Southern California, Los Angeles

γ-Herpesviruses (γ-HVs) establish life-long persistency in their host. Infection of mice with γ-HV68 provides a genetically tractable in vivo model for the characterization of the lifecycle/pathogenesis of γHVs. This protocol describes the detection and quantitation of γHV68 infection at acute and latent stages following infection by plaque-forming, infectious center, and qPCR assays.

Immunology and Infection

A Tetracycline-regulated Cell Line Produces High-titer Lentiviral Vectors that Specifically Target Dendritic Cells

Paul D. Bryson¹, Chupei Zhang¹, Chi-Lin Lee¹, Pin Wang¹

¹Mork Family Department of Chemical Engineering and Materials Science, Viterbi School of Engineering, University of Southern California, Los Angeles

Here, we use retroviral transduction and concatemeric transfection to create a cell line that can express the components of a lentiviral vector (LV) in the absence of tetracycline. This LV encodes GFP and is pseudotyped with a glycoprotein, SVGmu, which is specific for a receptor on dendritic cells.

Immunology and Infection

Laminar Flow-based Assays to Investigate Leukocyte Recruitment on Cultured Vascular Cells and Adherent Platelets

Tanja Vajen¹, Alexandra C.A. Heinzmann¹, Annemiek Dickhout¹, Zhen Zhao², Magdolna Nagy¹, Johan W.M. Heemskerk¹, Rory R. Koenen¹

¹Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, ²Department of Vascular Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine

Leukocytes avidly interact with vascular cells and platelets after vessel wall injury or during inflammation. Here, we describe a straightforward laminar flow-based assay to characterize the molecular mechanisms that underlie the interactions between leukocytes and their cellular partners.

Developmental Biology

Fast and Efficient Expression of Multiple Proteins in Avian Embryos Using mRNA Electroporation

Martin Tran¹, Mohit Dave^2,3, Rusty Lansford^1,2,3

¹Department of Biological Sciences, University of Southern California, Los Angeles, ²Department of Radiology and Developmental Neuroscience Program, Saban Research Institute, Children's Hospital Los Angeles, ³Keck School of Medicine, Department of Radiology, University of Southern California, Los Angeles

We report messenger RNA (mRNA) electroporation as a method that permits fast and efficient expression of multiple proteins in the quail embryo model system. This method can be used to fluorescently label cells and record their in vivo movements by time-lapse microscopy shortly after electroporation.

Behavior

Temporomandibular Joint Pain Measurement by Bite Force and Von Frey Filament Assays in Mice

Supawadee Jariyasakulroj^1,2, Qing Chang¹, Pao-Fen Ko¹, Yang Shu¹, Ziying Lin¹, Jingyi Chen¹, Jian-Fu Chen¹

¹Center for Craniofacial Molecular Biology, University of Southern California, Los Angeles, ²Department of Masticatory Science, Faculty of Dentistry, Mahidol University

This protocol describes how measuring bite force and head withdrawal threshold can capture pain behavior in Complete Freund's Adjuvant (CFA)-induced inflammatory temporomandibular joint (TMJ) pain.