University of Western Ontario 11 articles published in JoVE Neuroscience Evaluation of Synaptic Multiplicity Using Whole-cell Patch-clamp Electrophysiology Julia K. Sunstrum1, Wataru Inoue1,2,3 1Neuroscience Program, Schulich School of Medicine and Dentistry, University of Western Ontario, 2Robarts Research Institute, Schulich School of Medicine and Dentistry, University of Western Ontario, 3Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, University of Western Ontario Here, we present a protocol for evaluating the functional synaptic multiplicity using whole-cell patch clamp electrophysiology in acute brain slices. Biology Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast Yuwei Jiang1, Sonja Di Gregorio2, Maram B. Albakri1, Martin L. Duennwald1,2, Patrick Lajoie1 1Department of Anatomy and Cell Biology, University of Western Ontario, 2Department of Pathology and Laboratory Medicine, University of Western Ontario This article describes protocols to assess the effect of fluorescent proteins on the aggregation and toxicity of misfolded polyglutamine expansion for the rapid evaluation of a newly uncharacterized fluorescent protein in the context of fluorescent reporters. Neuroscience Conditions Affecting Social Space in Drosophila melanogaster Alison R. McNeil1, Sam N. Jolley1, Adesanya A. Akinleye2, Marat Nurilov2, Zulekha Rouzyi2, Austin J. Milunovich3, Moria C. Chambers3, Anne F. Simon1 1Department of Biology, University of Western Ontario, 2Department of Biology, York College/CUNY, 3Department of Entomology, Cornell University The effect of genes and environment on social space of Drosophila melanogaster can be quantified through a powerful but straightforward analytical paradigm. We show here different factors that can affect this social space, and thus need to be taken into consideration when designing experiments in this paradigm. Bioengineering Engineering 3D Cellularized Collagen Gels for Vascular Tissue Regeneration Sébastien Meghezi1, Dawit G. Seifu1,2, Nina Bono1,3, Larry Unsworth2,4,5, Kibret Mequanint6, Diego Mantovani1,2 1Laboratory for Biomaterials and Bioengineering, Department Min-Met-Materials Eng & CHU de Québec Research Center, Canada Research Chair I for the Innovation in Surgery, Laval University, 2NSERC CREATE Program for Regenerative Medicine (NCPRM), Laval University, 3Department Electronics, Information and Bioengineering, Politecnico di Milano, 4Department of Chemical and Materials Engineering, University of Alberta, 5National Institute for Nanotechnology, National Research Council (Canada), 6Department of Chemical and Biochemical Engineering, University of Western Ontario In this work, we present a technique for the rapid fabrication of living vascular tissues by direct culturing of collagen, smooth muscle cells and endothelial cells. In addition, a new protocol for the mechanical characterization of engineered vascular tissues is described. Developmental Biology Understanding Early Organogenesis Using a Simplified In Situ Hybridization Protocol in Xenopus Steven J. Deimling1, Rami R. Halabi2,3, Stephanie A. Grover4, Jean H. Wang2,5, Thomas A. Drysdale2,4,5 1Developmental and Stem Cell Biology, Hospital for Sick Children, 2Children's Health Research Institute, University of Western Ontario, 3Department of Physiology and Pharmacology, University of Western Ontario, 4Neurosciences and Mental Health, Hospital for Sick Children, 5Department of Paediatrics, University of Western Ontario The Xenopus laevis embryo continues to be exceptionally useful in the study of early development due to its large size and ease of manipulation. A simplified protocol for whole mount in situ hybridization protocol is provided that can be used in the identification of specific organs in this model system. Neuroscience Straightforward Assay for Quantification of Social Avoidance in Drosophila melanogaster Robert W. Fernandez1, Marat Nurilov2, Omar Feliciano2, Ian S. McDonald3, Anne F. Simon3 1Department of Molecular Biophysics and Biochemistry, Yale University, 2Department of Biology, York College/CUNY, 3Department of Biology, Western Ontario University Here, we present a protocol to quantify the avoidance of stressed individuals. This paradigm is powerful yet user-friendly and can be used to assess the influence of genes and environment on one kind of social interaction in Drosophila melanogaster. Environment Application of Two-spotted Spider Mite Tetranychus urticae for Plant-pest Interaction Studies Marc Cazaux*1,2, Marie Navarro*1,2, Kristie A. Bruinsma*1, Vladimir Zhurov*1, Tara Negrave1, Thomas Van Leeuwen3,4, Vojislava Grbic1,2, Miodrag Grbic1,2 1Department of Biology, The University of Western Ontario, 2Instituto de Ciencias de la Vid y el Vino, 3Department of Crop Protection, Ghent University, 4Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam Protocols for efficient preparation of homogenous samples of spider mites, infestation of experimental plants, and assessment of plant damage, as required for studies of plant-pest interaction were developed. Medicine Quantification of Breast Cancer Cell Invasiveness Using a Three-dimensional (3D) Model Donna Cvetković*1, Cameron Glenn-Franklin Goertzen*1, Moshmi Bhattacharya1,2,3 1Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, University of Western Ontario, 2Department of Oncology, Schulich School of Medicine and Dentistry, University of Western Ontario, 3Lawson Health Research Institute This article provides detailed methodologies for the use of three-dimensional (3D) assays to quantify breast cancer cell invasion. Specifically, we discuss the procedures required to set up such assays, quantification, and data analysis, as well as methods to examine the loss of membrane integrity that occurs when cells invade. Behavior A Method for Investigating Age-related Differences in the Functional Connectivity of Cognitive Control Networks Associated with Dimensional Change Card Sort Performance Bianca DeBenedictis1, J. Bruce Morton1 1Cognitive Development and Neuroimaging Laboratory, Department of Psychology, Brain and Mind Institute, University of Western Ontario This video presents a method of examining age-related changes in functional connectivity of cognitive control networks engaged by targeted tasks/processes. The technique is based on multi-variate analysis of fMRI data. Neuroscience Functional Imaging of Auditory Cortex in Adult Cats using High-field fMRI Trecia A. Brown1,6, Joseph S. Gati5, Sarah M. Hughes5, Pam L. Nixon1,6, Ravi S. Menon3,4,5, Stephen G. Lomber1,2,4,5,6,7 1Department of Physiology and Pharmacology, University of Western Ontario, 2Department of Psychology, University of Western Ontario, 3Department of Medical Biophysics, University of Western Ontario, 4Brain and Mind Institute, University of Western Ontario, 5Centre for Functional and Metabolic Mapping, Robarts Research Institute, University of Western Ontario, 6Cerebral Systems Laboratory, University of Western Ontario, 7National Centre for Audiology, University of Western Ontario Functional studies of the auditory system in mammals have traditionally been conducted using spatially-focused techniques such as electrophysiological recordings. The following protocol describes a method of visualizing large-scale patterns of evoked hemodynamic activity in the cat auditory cortex using functional magnetic resonance imaging. Biology Differentiation of Newborn Mouse Skin Derived Stem Cells into Germ-like Cells In vitro Paul William Dyce1,2 1Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, The University of Western Ontario, 2Children's Health Research Institute In recent years the formation of germ cells using in vitro culture methods has been demonstrated using several different types of somatic stem cells. This article will visually present the differentiation of newborn mouse derived stem cells to early oocyte-like cells.
