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Nihon University

9 ARTICLES PUBLISHED IN JoVE

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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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Environment

Protocol for Measuring the Thermal Properties of a Supercooled Synthetic Sand-water-gas-methane Hydrate Sample
Michihiro Muraoka 1, Naoko Susuki 1, Hiroko Yamaguchi 2, Tomoya Tsuji 3, Yoshitaka Yamamoto 1
1Research Institute of Energy Frontier, National Institute of Advanced Industrial Science and Technology (AIST), 2College of Industrial Technology, Nihon University, 3SHIZEN ikohza, Malaysia-Japan International Institute of Technology, Universiti Teknologi Malaysia

We present a protocol for measuring the thermal properties of synthetic hydrate-bearing sediment samples comprising sand, water, methane, and methane hydrate.

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Developmental Biology

An Efficient Method to Obtain Dedifferentiated Fat Cells
Hiroaki Taniguchi *1, Tomohiko Kazama *1, Kazuhiro Hagikura *1, Chii Yamamoto 1, Minako Kazama 1, Yuki Nagaoka 1, Taro Matsumoto 1
1Division of Cell Regeneration and Transplantation, School of Medicine, Nihon University

We have modified the conditions for DFAT cell generation and provide herein information regarding the use of an improved growth medium for the production of these cells.

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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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Biochemistry

Manipulating Living Cells to Construct Stable 3D Cellular Assembly Without Artificial Scaffold
Takehiro Yamazaki *1, Hiroaki Taniguchi *2, Shoto Tsuji 1, Shiho Sato 1, Takahiro Kenmotsu 1, Kenichi Yoshikawa 1, Koichiro Sadakane 1
1Faculty of Life and Medical Sciences, Doshisha University, 2The Institute of Genetics and Animal Breeding, Polish Academy of Sciences

We demonstrate a novel method for constructing a single-cell-based 3-dimensional (3D) assembly without an artificial scaffold.

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Developmental Biology

Neurogenesis Using P19 Embryonal Carcinoma Cells
Paweł Leszczyński 1, Magdalena Śmiech 1, Aamir S. Teeli 1, Aleksandra Zołocińska 2, Anna Słysz 2, Zygmunt Pojda 2, Mariusz Pierzchała 3, Hiroaki Taniguchi 1
1Department of Experimental Embryology, Institute of Genetics and Animal Breeding, Polish Academy of Sciences, 2Department of Regenerative Medicine, Maria Skłodowska-Curie Institute - Oncology Center, 3Department of Genomics and Biodiversities, Institute of Genetics and Animal Breeding, Polish Academy of Sciences

The P19 mouse embryonic carcinoma cell line (P19 cell line) is widely used for studying the molecular mechanism of neurogenesis with great simplification compared to in vivo analysis. Here, we present a protocol for retinoic acid-induced neurogenesis in the P19 cell line.

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Medicine

Novel Methods for Intranasal Administration Under Inhalation Anesthesia to Evaluate Nose-to-Brain Drug Delivery
Takanori Kanazawa *1, Mitsuyoshi Fukuda *1, Naoto Suzuki 1, Toyofumi Suzuki 1
1Laboratory of Pharmaceutics, School of Pharmacy, Nihon University

Here, we describe two novel methods of stable intranasal administration under inhalation anesthesia with minimal physical stress for experimental animals. We also describe a method for quantitative evaluation of drug distribution levels in the brain via the nose-to-brain pathway using radiolabeled [14C]-inulin as a model substrate of water-soluble macromolecules.

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Biochemistry

Quantitative Microtubule Fractionation Technique to Separate Stable Microtubules, Labile Microtubules, and Free Tubulin in Mouse Tissues
Ayaka Hagita-Tatsumoto 1,2, Tomohiro Miyasaka 1,2,3
1Department of Neuropathology, Faculty of Life and Medical Sciences, Doshisha University, 2Center for Research in Neurodegenerative Diseases, Doshisha University, 3Laboratory of Physiology and Anatomy, School of Pharmacy, Nihon University

Microtubules, which are tubulin polymers, play a crucial role as a cytoskeleton component in eukaryotic cells and are known for their dynamic instability. This study developed a method for fractionating microtubules to separate them into stable microtubules, labile microtubules, and free tubulin to evaluate the stability of microtubules in various mouse tissues.

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