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Collège de France

8 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents
Marie Vandecasteele 1,2, S. M. 1, Sébastien Royer 1,3, Mariano Belluscio 1, Antal Berényi 1, Kamran Diba 1,4, Shigeyoshi Fujisawa 1, Andres Grosmark 1, Dun Mao 1, Kenji Mizuseki 1, Jagdish Patel 1, Eran Stark 1, David Sullivan 1, Brendon Watson 1, György Buzsáki 1
1Center for Molecular and Behavioral Neuroscience, University of New Jersey, 2Center for Interdisciplinary Research in Biology, Collège de France, 3Janelia Farm Research Campus, Howards Hughes Medical Institute, 4Deptartment of Psychology, University of Wisconsin at Milwaukee

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

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Neuroscience

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices
Ulrike Pannasch *1, Jérémie Sibille *1,2, Nathalie Rouach 1
1Neuroglial Interactions in Cerebral Physiopathology, Center for Interdisciplinary Research in Biology, CNRS UMR 7241, INSERM U1050, Collège de France, 2Paris Diderot University

The preparation of acute brain slices from isolated hippocampi, as well as the simultaneous electrophysiological recordings of astrocytes and neurons in stratum radiatum during stimulation of schaffer collaterals is described. The pharmacological isolation of astroglial potassium and glutamate transporter currents is demonstrated.

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Medicine

Multi-electrode Array Recordings of Human Epileptic Postoperative Cortical Tissue
Elena Dossi 1, Thomas Blauwblomme 2,3, Rima Nabbout 2,4, Gilles Huberfeld 2,5, Nathalie Rouach 1
1Neuroglial Interactions in Cerebral Physiopathology, Center for Interdisciplinary Research in Biology, CNRS UMR 7241, INSERM U1050, Collège de France, 2Infantile Epilepsies & Brain Plasticity, INSERM U1129, PRES, Paris Descartes University, Sorbonne Paris Cité, CEA, 3Neurosurgery Department, Necker Hospital, AP-HP, Paris Descartes University, 4Rare Epilepsies Reference Center, Necker Hospital, AP-HP, Paris Descartes University, 5Neurophysiology Department, La Pitié-Salpêtrière Hospital, AP-HP, Sorbonne and Pierre and Marie Curie University

We here describe how to perform multi-electrode array recordings of human epileptic cortical tissue. Epileptic tissue resection, slice preparation and multi-electrode array recordings of interictal and ictal events are demonstrated in detail.

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Developmental Biology

Transcriptional Analysis by Nascent RNA FISH of In Vivo Trophoblast Giant Cells or In Vitro Short-term Cultures of Ectoplacental Cone Explants
Catherine Corbel 1, Edith Heard 1
1Unité de Génétique et Biologie du Développement, Institut Curie, PSL Research University, CNRS UMR 3215, INSERM U934

Trophoblast giant cells (TGCs) play a key role in the placenta to ensure a healthy pregnancy. We present a protocol for assessing the transcriptional status of genes in TGCs by nascent fluorescent in situ hybridization on cryostat sections of post-implantation embryos or short-term cultures of embryonic day 7 ectoplacental cones.

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Immunology and Infection

Immunofluorescence Analysis of Stress Granule Formation After Bacterial Challenge of Mammalian Cells
Pascale Vonaesch 1, Philippe J. Sansonetti 1,2, Pamela Schnupf 3
1Unité de Pathogénie Microbienne Moléculaire, INSERM U786, Institut Pasteur, 2Microbiologie et Maladies Infectieuses, Collège de France, 3Laboratory of Intestinal Immunity, Institut Imagine-INSERM UMR 1163, Université Paris Descartes-Sorbonne Paris Cité

We describe a method for the qualitative and quantitative analysis of stress granule formation in mammalian cells after the cells are challenged with bacteria and a number of different stresses. This protocol can be applied to investigate the cellular stress granule response in a wide range of host-bacterial interactions.

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Immunology and Infection

Imaging Ca2+ Responses During Shigella Infection of Epithelial Cells
Yasmine Smail 1,2,3,4, Chunhui Sun 1,2,3,4, Laurent Combettes 5, Guy Tran Van Nhieu 1,2,3,4
1Equipe Communication Intercellulaire et Infections Microbiennes, Centre de Recherche Interdisciplinaire en Biologie (CIRB), Collège de France, 2Institut National de la Santé et de la Recherche Médicale (Inserm), U1050, 3Centre Nationale de la Recherche Scientifique (CNRS), UMR7241, 4MEMOLIFE Laboratory of Excellence and Paris Sciences et Lettres, 5Inserm, UMRS1174, Université Paris Sud

Here, we present protocols to visualize calcium (Ca2+) responses elicited by HeLa cells infected by Shigella. By optimizing the parameters of bacterial infection and imaging with Ca2+ fluorescent probes, atypical global and local Ca2+ signals induced by bacteria over a large range of infection kinetics are characterized.

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Biology

Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe
Aline Stedman 1,3, Antonin Levy 1,4, Philippe J. Sansonetti 1,2,5, Giulia Nigro 1,6
1Molecular Microbial Pathogenesis Unit, Institut Pasteur, 2Chaire de Microbiologie et Maladies Infectieuses, Collège de France, 3Institut de Biologie Paris Seine (IBPS) - Developmental Biology Unit, Sorbonne Université, CNRS UMR7622, INSERM U1156, 4Molecular Radiotherapy, INSERM U1030, Gustave Roussy, Université Paris-Saclay, 5The Center for Microbes, Development and Health, Institut Pasteur Shanghai and Chinese Academy of Sciences, 6Microenvironment and Immunity Unit, Institut Pasteur

The present protocol describes a method to detect reactive oxygen species (ROS) in the intestinal murine organoids using qualitative imaging and quantitative cytometry assays. This work can be potentially extended to other fluorescent probes to test the effect of selected compounds on ROS.

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Biology

Deciphering High-Resolution 3D Chromatin Organization via Capture Hi-C
Antonia Hauth 1, Rafael Galupa 1, Nicolas Servant 2, Laura Villacorta 1, Kai Hauschulz 3, Joke Gerarda van Bemmel 4, Agnese Loda 1, Edith Heard 1,5
1EMBL: European Molecular Biology Laboratory, 2Institut Curie, 3Agilent Technologies, 4Genmab BV, 5Collège de France

This protocol describes the Capture Hi-C method used to characterize the 3D organization of megabased-sized targeted genomic regions at high-resolution, including boundaries of topologically associating domains (TADs) and long-range chromatin interactions between regulatory and other DNA sequence elements.

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