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MRC National Institute for Medical Research

4 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation
Emilie Pacary 1, Matilda A. Haas 1, Hendrik Wildner 1, Roberta Azzarelli 1, Donald M. Bell 2, Djoher Nora Abrous 3, François Guillemot 1
1Division of Molecular Neurobiology, MRC National Institute for Medical Research, 2Confocal and Image Analysis Laboratory, National Institute for Medical Research, 3Physiopathologie de la plasticité neuronale, Neurocentre Magendie, Université de Bordeaux

This article describes in detail a protocol to electroporate in utero the cerebral cortex and the hippocampus at E14.5 in mice. We also show that this is a valuable method to study dendrites and spines in these two cerebral regions.

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Developmental Biology

Genome-wide Snapshot of Chromatin Regulators and States in Xenopus Embryos by ChIP-Seq
George E. Gentsch 1, Ilya Patrushev 1, James C. Smith 1
1Division of Systems Biology, MRC National Institute for Medical Research

The question of how chromatin regulators and chromatin states affect the genome in vivo is key to our understanding of how early cell fate decisions are made in the developing embryo. ChIP-Seq—the most popular approach to investigate chromatin features at a global level—is outlined here for Xenopus embryos.

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Neuroscience

Adeno-associated Virus-mediated Transgene Expression in Genetically Defined Neurons of the Spinal Cord
Karen Haenraets *1,2, Gioele W. Albisetti *1, Edmund Foster 1, Hendrik Wildner 1
1Institute of Pharmacology and Toxicology, University of Zurich, 2Institute of Pharmaceutical Sciences, Swiss Federal Institute (ETH) Zurich

Intraspinal injection of recombinase dependent recombinant adeno-associated virus (rAAV) can be used to manipulate any genetically labelled cell type in the spinal cord. Here we describe how to transduce neurons in the dorsal horn of the lumbar spinal cord. This technique enables functional interrogation of the manipulated neuron subtype.

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Neuroscience

Fluorescence-Activated Nuclei Negative Sorting of Neurons Combined with Single Nuclei RNA Sequencing to Study the Hippocampal Neurogenic Niche
Thomas Kerloch 1, Tjaša Lepko 2, Kirill Shkura 2, François Guillemot 1, Sébastien Gillotin 2
1The Francis Crick Institute, 2MSD R&D Innovation Centre

Presented here is a method to sequence single nuclei isolated from the mouse dentate gyrus that excludes most neurons through fluorescence-activated nuclei (FAN)-sorting. This approach generates high-quality expression profiles and facilitates the study of most other cell types represented in the niche, including scarce populations such as neural stem cells.

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