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Sorbonne Université

7 ARTICLES PUBLISHED IN JoVE

Neuroscience

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

Gabrielle Devienne^*1, Benjamin Le Gac^*1, Juliette Piquet^*1, Bruno Cauli¹

¹UPMC Univ Paris 06, INSERM, CNRS, Neuroscience Paris Seine - Institut de Biologie Paris Seine (NPS - IBPS), Sorbonne Université

This protocol describes the critical steps and precautions required to perform single cell multiplex reverse transcription polymerase chain reaction after patch-clamp. This technique is a simple and effective method to analyze the expression profile of a predetermined set of genes from a single cell characterized by patch-clamp recordings.

Developmental Biology

Defining the Program of Maternal mRNA Translation during In vitro Maturation using a Single Oocyte Reporter Assay

Natasja G. J. Costermans^1,2, Enrico M. Daldello^1,2,3, Ria J. Marathe^1,2, Marco Conti^1,2

¹Center for Reproductive Sciences, University of California at San Francisco, ²Department of Obstetrics Gynecology and Reproductive Sciences, University of California at San Francisco, ³Present affiliation: Laboratoire de Biologie du Développement-Institut de Biologie Paris Seine, LBD-IBPS, Sorbonne Université

This protocol describes a reporter assay to study the regulation of mRNA translation in single oocytes during in vitro maturation.

Chemistry

Thermochemical Studies of Ni(II) and Zn(II) Ternary Complexes Using Ion Mobility-Mass Spectrometry

Anna J. Corrales¹, Anna V. Arredondo¹, Amber A. Flores¹, Chloe L. Duvak¹, Charles L. Mitchell¹, Riccardo Spezia², Laurence A. Angel¹

¹Department of Chemistry, Texas A&M University-Commerce, ²Laboratoire de Chimie Théorique, Sorbonne Université

This article describes an experimental protocol using electrospray-ion mobility-mass spectrometry, semi-empirical quantum calculations, and energy-resolved threshold collision-induced dissociation to measure the relative thermochemistry of the dissociation of related ternary metal complexes.

Biochemistry

Purification and Quality Control of Recombinant Septin Complexes for Cell-Free Reconstitution

Gerard Castro-Linares¹, Jeffrey den Haan¹, Francois Iv², Carla Silva Martins², Aurélie Bertin³, Manos Mavrakis², Gijsje H. Koenderink¹

¹Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, ²Institut Fresnel, CNRS UMR7249, Aix Marseille Univ, Centrale Marseille, ³Institut Curie, Université PSL, Sorbonne Université

In vitro reconstitution of cytoskeletal proteins is a vital tool to understand the basic functional properties of these proteins. The present paper describes a protocol to purify and assess the quality of recombinant septin complexes, which play a central role in cell division and migration.

Biology

Bottom-Up In Vitro Methods to Assay the Ultrastructural Organization, Membrane Reshaping, and Curvature Sensitivity Behavior of Septins

Brieuc Chauvin^*1, Koyomi Nakazawa^*1, Alexandre Beber^1,7, Aurélie Di Cicco¹, Bassam Hajj¹, François Iv², Manos Mavrakis², Gijsje H. Koenderink³, João T. Cabral⁴, Michaël Trichet⁵, Stéphanie Mangenot^*6, Aurélie Bertin^*1

¹Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University, Sorbonne Université, ²Institut Fresnel, CNRS UMR7249, Aix Marseille Univ, Centrale Marseille, ³Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, ⁴Department of Chemical Engineering, Imperial College London, ⁵Sorbonne Université, CNRS, Institut de Biologie Paris-Seine (IBPS), Service de microscopie électronique (IBPS-SME), ⁶Laboratoire Matière et Systèmes Complexes (MSC), Université Paris Cité, ⁷Institute of Biotechnology, Czech Academy of Sciences, BIOCEV

Septins are cytoskeletal proteins. They interact with lipid membranes and can sense but also generate membrane curvature at the micron scale. We describe in this protocol bottom-up in vitro methodologies for analyzing membrane deformations, curvature-sensitive septin binding, and septin filament ultrastructure.

Medicine

Robot-Assisted Transcanal Endoscopic Ear Surgery for Congenital Cholesteatoma

François Simon^1,2, Yann Nguyen^3,4, Natalie Loundon², Françoise Denoyelle^1,2

¹Université Paris Cité, ²Pediatric Otolaryngology, Head and neck surgery department, AP-HP, Hôpital Necker-Enfants Malades, ³Sorbonne Université, ⁴Otolaryngology - head and neck surgery department AP-HP, Hôpital Pitié-Salpêtrière

This protocol describes the removal of congenital cholesteatoma using minimally invasive transcanal endoscopic ear surgery with a two-handed approach and a robotic endoscope holder.

Developmental Biology

Efficient Vascularization of Kidney Organoids through Intracelomic Transplantation in Chicken Embryos

Marije Koning^1,2, Ellen Lievers^1,2, Thierry Jaffredo³, Cathelijne W. van den Berg^1,2,4, Ton J. Rabelink^1,2,4

¹Department of Internal Medicine - Nephrology, Leiden University Medical Center, ²Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center, ³IBPS, CNRS UMR7622, Developmental Biology Laboratory, Sorbonne Université, ⁴The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center

Here, we present a detailed protocol for the transplantation of kidney organoids in the celomic cavity of chicken embryos. This method induces vascularization and enhanced maturation of the organoids within 8 days and can be used to study these processes in an efficient manner.