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Université Laval

16 ARTICLES PUBLISHED IN JoVE

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Biology

Genome-wide Protein-protein Interaction Screening by Protein-fragment Complementation Assay (PCA) in Living Cells
Samuel Rochette *1, Guillaume Diss *1, Marie Filteau 1, Jean-Baptiste Leducq 1, Alexandre K. Dubé 1, Christian R. Landry 1
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).

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Biology

RNA Isolation from Cell Specific Subpopulations Using Laser-capture Microdissection Combined with Rapid Immunolabeling
Audrey Chabrat *1,2, Hélène Doucet-Beaupré *1,2, Martin Lévesque 1,2
1Department of Psychiatry and Neurosciences, Faculty of Medicine, Université Laval, 2Centre de recherche de l'Institut universitaire en santé mentale de Québec

Gene expression analysis of a subset of cells in a specific tissue represents a major challenge. This article describes how to isolate high-quality total RNA from a specific cell population by combining a rapid immunolabeling method with laser capture microdissection.

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JoVE Core

Automation of Mode Locking in a Nonlinear Polarization Rotation Fiber Laser through Output Polarization Measurements
Michel Olivier 1,2, Marc-Daniel Gagnon 1, Joé Habel 1
1Centre d'optique, photonique et laser, Université Laval, 2Département de physique, Cégep Garneau

A protocol to detect and automate mode locking in a pre-adjusted nonlinear polarization rotation fiber laser is presented. The detection of a sudden change in the output polarization state when mode locking occurs is used to command the alignment of an intra-cavity polarization controller in order to find mode-locking conditions.

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Developmental Biology

Transcriptome Profiling of In-Vivo Produced Bovine Pre-implantation Embryos Using Two-color Microarray Platform
Reza Salehi *1, Stephen C.M. Tsoi *1, Marcos G. Colazo 2, Divakar J. Ambrose 1,2, Claude Robert 3, Michael K. Dyck 1
1Department of Agricultural, Food and Nutritional Science, University of Alberta, 2Livestock Research Branch, Alberta Agriculture and Forestry, 3Laboratory of Functional Genomics of Early Embryonic Development, Université Laval

Microarray technology allows quantitative measurement and gene expression profiling of transcripts on a genome-wide basis. Therefore, this protocol provides an optimized technical procedure in a two-color custom made bovine array using Day 7 bovine embryos to demonstrate the feasibility of using low amount of total RNA.

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Neuroscience

Correlative Light and Electron Microscopy to Study Microglial Interactions with β-Amyloid Plaques
Kanchan Bisht *1, Hassan El Hajj *1, Julie C. Savage 1, Maria G. Sánchez 1, Marie-Ève Tremblay 1
1Neurosciences Axis, CHU de Québec Research Center

This article describes a protocol for visualizing amyloid Aβ plaques in Alzheimer's disease mouse models using methoxy-X04, which crosses the blood-brain barrier and selectively binds to β-pleated sheets found in dense core Aβ plaques. It allows pre-screening of plaque-containing tissue sections prior to immunostaining and processing for electron microscopy.

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Biology

Adenofection: A Method for Studying the Role of Molecular Chaperones in Cellular Morphodynamics by Depletion-Rescue Experiments
Margit Fuchs 1,2, Marie-Chloé Boulanger 3,4, Herman Lambert 1,2, Jacques Landry 1,2, Josée N. Lavoie 1,2
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.

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Neuroscience

Preparation of Non-human Primate Brain Tissue for Pre-embedding Immunohistochemistry and Electron Microscopy
Lara Eid 1,2, Martin Parent 1
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.

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Biochemistry

Quantitative Immunofluorescence to Measure Global Localized Translation
Jonathan Bergeman 1, Marc-Étienne Huot 1,2
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.

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Neuroscience

Two-photon Calcium Imaging in Neuronal Dendrites in Brain Slices
Olivier Camiré 1,2, Lisa Topolnik 1,2
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.

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Neuroscience

Ex Utero Electroporation and Organotypic Slice Cultures of Embryonic Mouse Brains for Live-Imaging of Migrating GABAergic Interneurons
Lara Eid 1,2, Mathieu Lachance 1, Gilles Hickson 1,3, Elsa Rossignol 1,2,4
1Centre de recherche du CHU Sainte-Justine, 2Department of Neuroscience, Université de Montréal, 3Department of pathology and cellular biology, Université de Montréal, 4Department of Pediatrics, Université de Montréal

Here, we provide a low-cost and reliable method to generate electroporated brain organotypic slice cultures from mouse embryos suitable for confocal microscopy and live-imaging techniques.

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Medicine

Assessment of Respiratory Function in Conscious Mice by Double-chamber Plethysmography
Samuel Mailhot-Larouche 1, Louis Deschênes 1, Katherine Lortie 1, Morgan Gazzola 1, David Marsolais 1, David Brunet 2, Annette Robichaud 2, Ynuk Bossé 1
1Institut Universitaire de Cardiologie et de Pneumologie de Québec, Université Laval, 2SCIREQ Scientific Respiratory Equipment Inc.

The objective of the present article is to provide a detailed description of the recommended procedures to evaluate respiratory function in conscious mice by double-chamber plethysmography.

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Neuroscience

Multi-Fiber Photometry to Record Neural Activity in Freely-Moving Animals
Ekaterina Martianova 1, Sage Aronson 2,3, Christophe D. Proulx 1
1CERVO Brain Research Center, Department of Psychiatry and Neurosciences, Université Laval, 2Center for Neural Circuits and Behavior, Department of Neuroscience and Section of Neurobiology, Division of Biology, University of California at San Diego, 3Neurophotometrics Ltd.

This protocol details how to implement and perform multi-fiber photometry recordings, how to correct for calcium-independent artifacts, and important considerations for dual-color photometry imaging.

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Immunology and Infection

Immunofluorescence Staining Using IBA1 and TMEM119 for Microglial Density, Morphology and Peripheral Myeloid Cell Infiltration Analysis in Mouse Brain
Fernando González Ibanez 1,2, Katherine Picard 1,2, Maude Bordeleau 1,3, Kaushik Sharma 1,2,4, Kanchan Bisht 1,2,4, Marie-Ève Tremblay 1,2
1Axe Neurosciences, Centre de Recherche du CHU de Québec-Université Laval, 2Département de Médecine Moléculaire, Faculté de Médecine, Université Laval, 3Integrated Program in Neuroscience, McGill University, 4Center for Brain Immunology and Glia (BIG), University of Virginia

This protocol describes a step-by-step workflow for immunofluorescent costaining of IBA1 and TMEM119, in addition to analysis of microglial density, distribution, and morphology, as well as peripheral myeloid cell infiltration in mouse brain tissue.

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Immunology and Infection

Precise Brain Mapping to Perform Repetitive In Vivo Imaging of Neuro-Immune Dynamics in Mice
Kanchan Bisht 1,2, Kaushik Sharma 1,2, Ukpong B. Eyo 1,2
1Center for Brain Immunology and Glia (BIG), University of Virginia, 2Department of Neuroscience, University of Virginia

This protocol describes a chronic cranial window implantation technique that can be used for longitudinal imaging of neuro-glio-vascular structures, interactions, and function in both healthy and diseased conditions. It serves as a complementary alternative to the transcranial imaging approach that, while often preferred, possesses some critical limitations.

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Biology

Evaluation of the Efficacy of Organic Peroxyacids for Eradicating Dairy Biofilms Using an Approach Combining Static and Dynamic Methods
Coralie Goetz *1, Nissa Niboucha *1, Eric Jubinville 1, Julie Jean 1
1Département des Sciences des Aliments, Institut sur la Nutrition et les Aliments Fonctionnels, Université Laval

This protocol describes an approach combining static and dynamic methods to evaluate the efficacy of organic peroxyacids for eradicating biofilms in the dairy industry. This approach may also be used to test the effectiveness of new biological or chemical formulations for controlling biofilms.

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Environment

Inoculating and Observing Arbuscular Mycorrhizal Cultures on Superabsorbent Polymer-Based Autotrophic Systems
Louis Paré 1, Claudia Banchini 2, Franck Stefani 2
1Centre d’Étude de la Forêt (CEF) and Institut de Biologie Intégrative et des Systèmes (IBIS), Université Laval, 2Ottawa Research and Development Centre, Agriculture and Agri-Food Canada

Here, we describe a simple and inexpensive technique for inoculating and observing arbuscular mycorrhizal fungi in superabsorbent polymer-based autotrophic systems.

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