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PSL Research University

3 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Stem cell-like Xenopus Embryonic Explants to Study Early Neural Developmental Features In Vitro and In Vivo
Beatrice C. Durand 1,2,3,4
1Institut Curie, 2UMR 3387, CNRS, 3PSL Research University, 4Université Paris-Sud

In Xenopus embryos, cells from the roof of the blastocoel are pluripotent and can be programmed to generate various tissues. Here, we describe protocols to use amphibian blastocoel roof explants as an assay system to investigate key in vivo and in vitro features of early neural development.

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Medicine

Electroporation-Based Genetic Modification of Primary Human Pigment Epithelial Cells Using the Sleeping Beauty Transposon System
Sandra Johnen 1, Nina Harmening 2,3, Corinne Marie 4,5, Daniel Scherman 4, Zsuzsanna Izsvák 6, Zoltán Ivics 7, Peter Walter 1, Gabriele Thumann 2,3
1Department of Ophthalmology, University Hospital RWTH Aachen, 2Experimental Ophthalmology, University of Geneva, 3Department of Ophthalmology, University Hospitals of Geneva, 4Université de Paris, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, 5Chimie ParisTech, PSL Research University, 6Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 7Division of Medical Biotechnology, Paul-Ehrlich-Institute

We have developed a protocol to transfect primary human pigment epithelial cells by electroporation with the gene encoding pigment epithelium-derived factor (PEDF) using the Sleeping Beauty (SB) transposon system. Successful transfection was demonstrated by quantitative polymerase chain reaction (qPCR), immunoblotting, and enzyme-linked immunosorbent assay (ELISA).

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Neuroscience

Whole-Brain 3D Activation and Functional Connectivity Mapping in Mice using Transcranial Functional Ultrasound Imaging
Adrien Bertolo *1, Mohamed Nouhoum *1, Silvia Cazzanelli *1, Jeremy Ferrier *3, Jean-Charles Mariani 2, Andrea Kliewer 4,2, Benoit Belliard 1, Bruno-Félix Osmanski 3, Thomas Deffieux *1, Sophie Pezet *1, Zsolt Lenkei *2, Mickael Tanter *1
1Physics for Medicine Paris, ESPCI Paris, INSERM, CNRS, PSL Research University, 2Institute of Psychiatry and Neurosciences of Paris, INSERM U1266, Université de Paris, 3Iconeus, 4Department of Pharmacology and Toxicology, Jena University Hospital - Friedrich Schiller University Jena

This protocol describes the quantification of volumetric cerebral hemodynamic variations in the mouse brain using functional ultrasound (fUS). Procedures for 3D functional activation map following sensory stimulation as well as resting-state functional connectivity are provided as illustrative examples, in anesthetized and awake mice.

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