Collège de France

8 ARTICLES PUBLISHED IN JoVE

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 Belluscio¹, Antal Berényi¹, Kamran Diba^1,4, Shigeyoshi Fujisawa¹, Andres Grosmark¹, Dun Mao¹, Kenji Mizuseki¹, Jagdish Patel¹, Eran Stark¹, David Sullivan¹, Brendon Watson¹, György Buzsáki¹

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.

Neuroscience

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Ulrike Pannasch^*1, Jérémie Sibille^*1,2, Nathalie Rouach¹

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.

Medicine

Multi-electrode Array Recordings of Human Epileptic Postoperative Cortical Tissue

Elena Dossi¹, Thomas Blauwblomme^2,3, Rima Nabbout^2,4, Gilles Huberfeld^2,5, Nathalie Rouach¹

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.

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 Corbel¹, Edith Heard¹

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.

Immunology and Infection

Immunofluorescence Analysis of Stress Granule Formation After Bacterial Challenge of Mammalian Cells

Pascale Vonaesch¹, Philippe J. Sansonetti^1,2, Pamela Schnupf³

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.

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 Combettes⁵, 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.

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.

Biology

Deciphering High-Resolution 3D Chromatin Organization via Capture Hi-C

Antonia Hauth¹, Rafael Galupa¹, Nicolas Servant², Laura Villacorta¹, Kai Hauschulz³, Joke Gerarda van Bemmel⁴, Agnese Loda¹, 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.