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RIKEN Brain Science Institute

10 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection
Asuka Matsui 1, Aya C. Yoshida 1, Mayumi Kubota 1, Masaharu Ogawa 1, Tomomi Shimogori 1
1Lab for Molecular Mechanisms of Thalamus Development, RIKEN Brain Science Institute

A gene transfer method into the developing mouse brain is described by using a unique surgical method and special shape of electrodes. This unique technique allows transfection of plasmid DNA temporally and spatially, which will aid many neuroscientists in studying brain development.

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Neuroscience

Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics
M. Rezaul Karim 1,2, Adrian W. Moore 1
1Disease Mechanism Research Core, RIKEN Brain Science Institute, 2Graduate School of Science and Engineering, Saitama University

The dendritic arborization sensory neurons of the Drosophila larval peripheral nervous system are useful models to elucidate both general and neuron class-specific mechanisms of neuron differentiation. We present a practical guide to generate and analyze dendritic arborization neuron genetic mosaics.

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Neuroscience

Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall
Cagri Yalgin 1,2, M. Rezaul Karim 1,2, Adrian W. Moore 1
1Disease Mechanism Research Core, RIKEN Brain Science Institute, 2Graduate School of Science and Engineering, Saitama University

To understand how complex cell shapes, such as neuronal dendrites, are achieved during development, it is important to be able to accurately assay microtubule organization. Here we describe a robust immunohistological labeling method to examine microtubule organization of dendritic arborization neuron sensory dendrites, trachea, muscle, and other Drosophila larva body wall tissues.

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Neuroscience

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna
M. Rezaul Karim 1, Keita Endo 2, Adrian W Moore 3, Hiroaki Taniguchi 1
1Laboratory for Genetic Code, Graduate School of Life and Medical Sciences, Doshisha University, 2Laboratory for Circuit Mechanisms of Sensory Perception, RIKEN Brain Science Institute, 3Disease Mechanism Research Core, RIKEN Brain Science Institute

Herein we describe the process of whole mount immunostaining of Drosophila antennae, which enables us to better understand the molecular mechanisms involved in the diversification of olfactory receptor neurons (ORN)s.

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Developmental Biology

Imaging of Cell Shape Alteration and Cell Movement in Drosophila Gastrulation Using DE-cadherin Reporter Transgenic Flies
M. Rezaul Karim 1, Tomohiro Haruta 2, Taro Matsumoto 3, Hiroki Oda 2, Hiroaki Taniguchi 3,4,5
1Biotechnology and Genetic Engineering Department, Jahangirnagar University, 2JT Biohistory Research Hall, 3Division of Cell Regeneration and Transplantation, School of Medicine, Nihon University, 4Graduate School of Life and Medical Sciences, Doshisha University, 5Institute of Genetics and Animal Breeding of the Polish Academy of Sciences

Herein we describe a procedure to capture live images of Drosophila gastrulation. This has enabled us to better understand the apical constriction involved in early development and further analyze mechanisms governing cellular movements during tissue structure modification.

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Biochemistry

A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis
Megumi Tsuchiya *1, M. Rezaul Karim *2, Taro Matsumoto 3, Hidesato Ogawa 1, Hiroaki Taniguchi 3,4,5
1Graduate School of Frontier Biosciences, Osaka University, 2Department of Biotechnology and Genetic Engineering, Jahangirnagar University, 3Division of Cell Regeneration and Transplantation, School of Medicine, Nihon University, 4Institute of Genetics and Animal Breeding of the Polish Academy of Sciences, 5Graduate School of Life and Medical Sciences, Doshisha University

We have established a method for the purification of coregulatory interaction proteins using the LC-MS/MS system.

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Neuroscience

Large-scale Three-dimensional Imaging of Cellular Organization in the Mouse Neocortex
Taisuke Yoneda 1, Seiichiro Sakai 1,2, Hisato Maruoka 1, Toshihiko Hosoya 1
1RIKEN Brain Science Institute, 2Tokyo Metropolitan Institute of Medical Science

Here we describe a procedure for tissue clearing, fluorescent labeling, and large-scale imaging of mouse brain tissue which, thereby, enables visualization of the three-dimensional organization of cell types in the neocortex.

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Neuroscience

Purification of the Dendritic Filopodia-rich Fraction
Yutaka Furutani 1,2, Yoshihiro Yoshihara 3
1Laboratory for Neurobiology of Synapse, RIKEN Brain Science Institute, 2Liver Cancer Prevention Research Unit, RIKEN Center for Integrative Medical Sciences, 3Laboratory for Systems Molecular Ethology, RIKEN Center for Brain Science

In this protocol, we introduce a method for purifying the dendritic filopodia-rich fraction from the phagocytic cup-like protrusion structure on cultured hippocampal neurons by taking advantage of the specific and strong affinity between a dendritic filopodial adhesion molecule, TLCN, and an extracellular matrix molecule, vitronectin.

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Neuroscience

In Vivo Chronic Two-Photon Imaging of Microglia in the Mouse Hippocampus
Ryosuke Kamei 1, Shinji Urata 1, Hisato Maruoka 1, Shigeo Okabe 1
1Department of Cellular Neurobiology, Graduate School of Medicine, The University of Tokyo

This paper describes a method for chronic in vivo observation of the resting microglia in the mouse hippocampal CA1 using precisely controlled surgery and two-photon microscopy.

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Neuroscience

Simultaneous Imaging of Microglial Dynamics and Neuronal Activity in Awake Mice
Hisato Maruoka 1, Ryosuke Kamei 1, Shunsuke Mizutani 1, Qingrui Liu 1, Shigeo Okabe 1
1Department of Cellular Neurobiology, Graduate School of Medicine and Faculty of Medicine, The University of Tokyo

Here, we describe a protocol combining adeno-associated virus injection with cranial window implantation for simultaneous imaging of microglial dynamics and neuronal activity in awake mice.

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