Laval University View Institution's Website 12 articles published in JoVE Neuroscience Two-photon Calcium Imaging in Neuronal Dendrites in Brain Slices Olivier Camiré1,2, Lisa Topolnik1,2 1CHU de Québec-Université Laval Research Center, Université Laval, 2Department of Biochemistry, Microbiology and Bioinformatics, Université Laval We present a method combining whole-cell patch-clamp recordings and two-photon imaging to record Ca2+ transients in neuronal dendrites in acute brain slices. Biochemistry Quantitative Immunofluorescence to Measure Global Localized Translation Jonathan Bergeman1, Marc-Étienne Huot1,2 1Centre de Recherche sur le Cancer de l’Université Laval, Faculté de Médecine, Département de Biologie moléculaire, biochimie médicale et pathologie, Université Laval, 2CRCHU de Québec: L’Hôtel-Dieu de Québec This manuscript describes a method to visualize and quantify localized translation events in subcellular compartments. The approach proposed in this manuscript requires a basic confocal imaging system and reagents and is rapid and cost-effective. Neuroscience Preparation of Non-human Primate Brain Tissue for Pre-embedding Immunohistochemistry and Electron Microscopy Lara Eid1,2, Martin Parent1 1Centre de recherche de l'Institut universitaire en santé mentale de Québec, Department of Psychiatry and Neuroscience, Université Laval, 2Centre de recherche du CHU Sainte-Justine Here, we provide an easy, low-cost, and time-efficient protocol to chemically fix primate brain tissue with acrolein fixative, allowing for long-term preservation that is compatible with pre-embedding immunohistochemistry for transmission electron microscopy. Biology Adenofection: A Method for Studying the Role of Molecular Chaperones in Cellular Morphodynamics by Depletion-Rescue Experiments Margit Fuchs1,2, Marie-Chloé Boulanger3,4, Herman Lambert1,2, Jacques Landry1,2, Josée N. Lavoie1,2 1Département de biologie moléculaire, biochimie médicale et pathologie, Faculté de médecine, Centre de recherche sur le cancer de l'Université Laval, 2Oncology, Centre de recherche du CHU de Québec, Université Laval, 3Laboratoire d'études moléculaires des valvulopathies (LEMV), Groupe de recherche en valvulopathies (GRV), Quebec Heart and Lung Institute/Research Center, 4Department of Surgery, Université Laval We describe a method for depletion-rescue experiments that preserves cellular integrity and protein homeostasis. Adenofection enables functional analyses of proteins within biological processes that rely on finely tuned actin-based dynamics, such as mitotic cell division and myogenesis, at the single-cell level. Medicine Three-Dimensional Culture Assay to Explore Cancer Cell Invasiveness and Satellite Tumor Formation Marie-France Côté1, Audrey Turcotte1,2, Charles Doillon1,3, Stephane Gobeil1,2 1CHU de Québec Research Centre, 2Department of Molecular Medicine, Laval University, 3Department of Surgery, Laval University Cancer cells are embedded in a collagen gel and then sandwiched in an acellular fibrin gel to generate a 3D culture system in which the invasiveness and formation of satellite tumors may be monitored. Neuroscience Electrophysiology on Isolated Brainstem-spinal Cord Preparations from Newborn Rodents Allows Neural Respiratory Network Output Recording Jean-Philippe Rousseau1, Céline Caravagna1 1Department of Pediatrics, Laval University The central respiratory drive is located in the brainstem. Spontaneous respiratory motor output from an isolated brainstem-spinal cord is recorded by placing an electrode on the fourth ventral root. This experimental approach is valuable for pharmacological investigations or the assessment of respiratory challenges and genetic manipulations on rhythmic motor behavior. 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. Medicine Increasing Pulmonary Artery Pulsatile Flow Improves Hypoxic Pulmonary Hypertension in Piglets Audrey Courboulin1, Chantal Kang2, Olivier Baillard3,4,5, Sebastien Bonnet1, Pierre Bonnet6 1Department of Medicine, Pulmonary Hypertension Research Group (CRIUCPQ), Laval University, 2Institut National de la Recherche Agronomique, 3Université Diderot Paris, Sorbonne Paris Cité, 4Hôpital Lariboisière, Physiologie clinique Explorations Fonctionnelles, 5INSERM U 965, 6Service de Cardiologie, Centre Hospitalier Universitaire Tours Pulmonary hypertension is associated with a significant reduction in pulmonary artery pulsatility, contractility and elasticity, contributing to an increase in pulmonary artery pressure and pulmonary resistance. Using a hypoxic piglet model, this study demonstrated that improving pulmonary artery plasticity using a newly developed pulsatile catheter improves hypoxic pulmonary hypertension. Developmental Biology Generation of Human Adipose Stem Cells through Dedifferentiation of Mature Adipocytes in Ceiling Cultures Julie Lessard1, Julie Anne Côté1, Marc Lapointe1, Mélissa Pelletier2, Mélanie Nadeau1, Simon Marceau1, Laurent Biertho1, André Tchernof1,2,3 1IUCPQ Research Center, 2CHU de Québec Research Center, 3Laval University Mature adipocytes may represent an abundant source of stem cells through dedifferentiation, which leads to a homogenous population of fibroblast-like cells. Collagenase digestion is used to isolate mature adipocytes from human fat. The goal of our protocol is to obtain multipotent, dedifferentiated fat cells from human mature adipocytes. Biology Genome-wide Protein-protein Interaction Screening by Protein-fragment Complementation Assay (PCA) in Living Cells Samuel Rochette*1, Guillaume Diss*1, Marie Filteau1, Jean-Baptiste Leducq1, Alexandre K. Dubé1, Christian R. Landry1 1Département de Biologie, Institut de biologie intégrative et des systémes & PROTEO, Université Laval Proteins interact with each other and these interactions determine in a large part their functions. Protein interaction partners can be identified at high-throughput in vivo using a yeast fitness assay based on the dihydrofolate reductase protein-fragment complementation assay (DHFR-PCA). Biology Analysis of Translation Initiation During Stress Conditions by Polysome Profiling Laëtitia Coudert*1,2, Pauline Adjibade*1,2, Rachid Mazroui1,2 1Department of Molecular Biology, Medical Biochemistry, and Pathology, Faculty of Medicine, Laval University, 2CHU de Quebec Research Center Here, we describe a method to analyze changes in the initiation of mRNA translation of eukaryotic cells in response to stress conditions. This method is based on the velocity separation on sucrose gradients of translating ribosomes from non-translating ribosomes. Medicine Mouse Model of Intraluminal MCAO: Cerebral Infarct Evaluation by Cresyl Violet Staining Estelle Rousselet1, Jasna Kriz2, Nabil G. Seidah1 1Laboratory of Biochemical Neuroendocrinology, Clinical Research Institute of Montreal, 2CHUQ Research Centre, Laval University The intraluminal middle cerebral occlusion model in mice is herein presented. The extent of cerebral infarct is evaluated by a neurologic score and cresyl violet staining, an alternative staining to TTC, offering the great advantage to test in parallel many interest markers.