University of Otago 4 articles published in JoVE Immunology and Infection Identifying Caspases and their Motifs that Cleave Proteins During Influenza A Virus Infection Matloob Husain1 1Department of Microbiology and Immunology, University of Otago Influenza A virus (IAV) infection activates the caspases that cleave host and viral proteins, which, in turn, have pro- and antiviral functions. By employing inhibitors, RNA interference, site-directed mutagenesis, and western blotting and RT-qPCR techniques, caspases in infected mammalian cells that cleave host cortactin and histone deacetylases were identified. Biology Small-Scale Plasma Membrane Preparation for the Analysis of Candida albicans Cdr1-mGFPHis Golnoush Madani*1, Erwin Lamping*1, Hee Ji Lee1, Masakazu Niimi1,2, Alok K. Mitra3, Richard D. Cannon1 1Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, 2Department of Microbiology, Faculty of Medicine, Chulalongkorn University, 3School of Biological Sciences, University of Auckland This article presents a small-scale plasma membrane isolation protocol for the characterization of Candida albicans ABC (ATP-binding cassette) protein Cdr1, overexpressed in Saccharomyces cerevisiae. A protease-cleavable C-terminal mGFPHis double tag with a 16-residue linker between Cdr1 and the tag was designed to facilitate the purification and detergent-screening of Cdr1. Neuroscience A Time-Efficient Fluorescence Spectroscopy-Based Assay for Evaluating Actin Polymerization Status in Rodent and Human Brain Tissues Faraz Ahmad1, Ping Liu1 1Department of Anatomy, School of Biomedical Sciences, University of Otago We report a simple, time-efficient and high-throughput fluorescence spectroscopy-based assay for the quantification of actin filaments in ex vivo biological samples from brain tissues of rodents and human subjects. Neuroscience Organotypic Hippocampal Slice Cultures As a Model to Study Neuroprotection and Invasiveness of Tumor Cells Urszula Grabiec*1, Tim Hohmann*1, Niels Hammer2, Faramarz Dehghani1 1Department of Anatomy and Cell Biology, Martin Luther University Halle-Wittenberg, 2Department of Anatomy, University of Otago Organotypic hippocampal slice cultures (OHSC) represent an in vitro model that simulates the in vivo situation very well. Here we describe a vibratome-based improved slicing protocol to obtain high quality slices for use in assessing the neuroprotective potential of novel substances or the biological behavior of tumor cells.
