PSL Research University

3 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Stem cell-like Xenopus Embryonic Explants to Study Early Neural Developmental Features In Vitro and In Vivo

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.

Medicine

Electroporation-Based Genetic Modification of Primary Human Pigment Epithelial Cells Using the Sleeping Beauty Transposon System

Sandra Johnen¹, Nina Harmening^2,3, Corinne Marie^4,5, Daniel Scherman⁴, Zsuzsanna Izsvák⁶, Zoltán Ivics⁷, Peter Walter¹, 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).

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 Mariani², Andrea Kliewer^4,2, Benoit Belliard¹, Bruno-Félix Osmanski³, 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.