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Aix-Marseille University

10 ARTICLES PUBLISHED IN JoVE

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Biology

Mapping Molecular Diffusion in the Plasma Membrane by Multiple-Target Tracing (MTT)
Vincent Rouger 1,2,3, Nicolas Bertaux 4,5,6, Tomasz Trombik 1,2,3, Sébastien Mailfert 1,2,3, Cyrille Billaudeau 1,2,3, Didier Marguet 1,2,3, Arnauld Sergé 1,2,3
1Institut National de la Santé et de la Recherche Médicale, UMR 631, Parc scientifique de Luminy, 2Centre National de la Recherche Scientifique, UMR 6102, Parc scientifique de Luminy, 3Centre d'Immunologie de Marseille-Luminy, Aix-Marseille University, 4École Centrale Marseille, Technopôle de Château-Gombert, 5Institut Fresnel, Aix-Marseille University, 6Centre National de la Recherche Scientifique, UMR 6133, Aix-Marseille University

Multiple-Target Tracing is a homemade algorithm developed for tracking individually labeled molecules within the plasma membrane of living cells. Efficiently detecting, estimating and tracing molecules over time at high-density provide a user-friendly, comprehensive tool to investigate nanoscale membrane dynamics.

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Biology

Cell Labeling and Injection in Developing Embryonic Mouse Hearts
Emilye Hiriart *1, Patrick van Vliet *2, Ralf J. Dirschinger 2, Sylvia M. Evans 2, Michel Puceat 1
1INSERM UMR-910, Aix-Marseille University, 2Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego

We describe a series of methods to inject dyes, DNA vectors, virus, and cells in order to monitor both cell fate and phenotype of endogenous and grafted cells derived from embryonic or pluripotent cells within mouse embryos at embryonic day (E)9.5 and later stages of development.

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JoVE Journal

Assessment of Social Cognition in Non-human Primates Using a Network of Computerized Automated Learning Device (ALDM) Test Systems
Joël Fagot 1, Yousri Marzouki 1, Pascal Huguet 1, Julie Gullstrand 1, Nicolas Claidière 1
1CNRS, Federation 3C, and Laboratory of Cognitive Psychology, Aix-Marseille University

Fagot & Paleressompoulle1 have published an automated learning device (ALDM) aimed at testing individual cognitive abilities in semi-free ranging monkeys. The main goal of our protocol is to use a network of ALDM test units to study social cognition in non-human primates.

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Neuroscience

A Novel Strategy Combining Array-CGH, Whole-exome Sequencing and In Utero Electroporation in Rodents to Identify Causative Genes for Brain Malformations
Valerio Conti *1, Aurelie Carabalona *2,3,15, Emilie Pallesi-Pocachard 2,3,4, Richard J. Leventer 5,6,7, Fabienne Schaller 2,3,8, Elena Parrini 1, Agathe A. Deparis 2,3, Françoise Watrin 2,3, Emmanuelle Buhler 2,3,8, Francesca Novara 9, Stefano Lise 10, Alistair T. Pagnamenta 10, Usha Kini 11, Jenny C. Taylor 10, Orsetta Zuffardi 9,12, Alfonso Represa 2,3, David Antony Keays 13, Renzo Guerrini 1,14, Antonio Falace 2,3, Carlos Cardoso 2,3
1University of Florence, 2INSERM INMED, 3Aix-Marseille University, 4Plateforme Biologie Moléculaire et Cellulaire INMED, 5Royal Children's Hospital, 6Murdoch Children's Research Institute, 7University of Melbourne, 8Plateforme postgenomique INMED, 9University of Pavia, 10Wellcome Trust Centre for Human Genetics, 11Oxford Radcliffe NHS Trust, 12IRCCS Casimiro Mondino Foundation, 13Research Institute of Molecular Pathology, 14IRCCS Stella Maris, 15Columbia University

Periventricular nodular heterotopia (PNH) is the most common form of malformation of cortical development (MCD) in adulthood but its genetic basis remains unknown in most sporadic cases. We have recently developed a strategy to identify novel candidate genes for MCDs and to directly confirm their causative role in vivo.

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Developmental Biology

Epigenetic Regulation of Cardiac Differentiation of Embryonic Stem Cells and Tissues
Imen Jebeniani 1, Julia Leschik 2, Michel Puceat 1
1INSERM UMR-910, Faculté de Médecine, Université Aix-Marseille, 2Institute of Physiological Chemistry, University Medical Center, Johannes Gutenberg University

A fine tuning regulation of gene transcription underlies embryonic cell fate decision. Herein, we describe chromatin immunoprecipitation assays used to investigate epigenetic regulation of both cardiac differentiation of stem cells and cardiac development of mouse embryos.

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Medicine

Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension
David Boulate 1, Myriam Amsallem 1,2, Jean-Baptiste Menager 1, Simon Dang Van 1, Peter Dorfmuller 3, Andrew Connolly 4, Alban Todesco 5, Benoit Decante 1, Elie Fadel 1, Francois Haddad 2, Olaf Mercier 1
1Research and Innovation Unit, RHU BioArt Lung 2020, Marie Lannelongue Hospital, 2Phenotypic and Biomarker Core Laboratory, Cardiovascular Institute, Stanford University, 3Department of pathology, Marie Lannelongue Hospital, 4Department of pathology, UCSF School of Medicine, 5Department of thoracic surgery, Hopital Nord, APHM, Aix-Marseille University

We present a protocol to induce and phenotype an acute right heart failure in a large animal model with chronic pulmonary hypertension. This model can be used to test therapeutic interventions, to develop right heart metrics or to improve the understanding of acute right heart failure pathophysiology.

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JoVE Journal

Highlighting and Reducing the Impact of Negative Aging Stereotypes During Older Adults' Cognitive Testing
Marie Mazerolle 1, Isabelle Régner 2, François Rigalleau 3, Pascal Huguet 4
1Department and Laboratory of Psychology, MSHE, Université Bourgogne Franche-Comté, 2Aix-Marseille University, CNRS, LPC, 3Department of Psychology, University of Poitiers, CNRS, 4University of Clermont Auvergne, CNRS, LAPSCO

Here, we present a protocol designed to show how negative aging stereotypes can impair memory performance of older adults during cognitive testing and how to reduce this deleterious effect. This method can help older people to perform at an optimal level during testing in both lab studies and clinical settings.

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Biochemistry

Spot Variation Fluorescence Correlation Spectroscopy for Analysis of Molecular Diffusion at the Plasma Membrane of Living Cells
Sebastien Mailfert *1, Karolina Wojtowicz *2, Sophie Brustlein *1, Ewa Blaszczak 3, Nicolas Bertaux 4, Marcin Łukaszewicz 2, Didier Marguet 1, Tomasz Trombik 2
1CNRS, Inserm, Centre d'Immunologie de Marseille-Luminy, Aix Marseille Univ, 2Faculty of Biotechnology, University of Wroclaw, 3Faculty of Biological Sciences, University of Wroclaw, 4CNRS, Centrale Marseille, Institut Fresnel, Aix Marseille Univ

This article aims to present a protocol on how to build a spot variation Fluorescence Correlation Spectroscopy (svFCS) microscope to measure molecular diffusion at the plasma membrane of living cells.

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Developmental Biology

Nuclei Isolation from Mouse Cardiac Progenitor Cells for Epigenome and Gene Expression Profiling at Single-Cell Resolution
Stephanie Ibrahim 1, Catherine Robert 2, Camille Humbert 2, Clotilde Ferreira 1, Gwenaëlle Collod 2, Sonia Stefanovic 1
1Center for Cardiovascular Disease & Nutrition, U1263, Aix-Marseille University, 2Marseille Medical Genetics, U1251, Aix-Marseille University

Here, we present a protocol describing cell nuclei preparation. After microdissection and enzymatic dissociation of cardiac tissue into single cells, the progenitor cells were frozen, followed by isolation of pure viable cells, which were used for single-nucleus RNA sequencing and the single-nucleus assay for transposase-accessible chromatin with high-throughput sequencing analyses.

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Neuroscience

Inducing Long-Term Plasticity of Intrinsic Neuronal Excitability in Neurons of the Dorsal Lateral Geniculate Nucleus
Michaël Russier 1, Maël Duménieu 1, Anushka Wakade 1, Salvatore Incontro 1, Laure Fronzaroli-Molinieres 1, Dominique Debanne 1
1Unité de Neurobiologie des canaux Ioniques et de la Synapse (UNIS), UMR1072, INSERM, Aix-Marseille University

Here, we describe a protocol for inducing long-term plasticity of neuronal intrinsic excitability in relay neurons from the dorsal lateral geniculate nucleus maintained in ex vivo brain slices.

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