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Nihon University

9 ARTICLES PUBLISHED IN JoVE

Neuroscience

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

M. Rezaul Karim¹, Keita Endo², Adrian W Moore³, Hiroaki Taniguchi¹

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.

Environment

Protocol for Measuring the Thermal Properties of a Supercooled Synthetic Sand-water-gas-methane Hydrate Sample

Michihiro Muraoka¹, Naoko Susuki¹, Hiroko Yamaguchi², Tomoya Tsuji³, Yoshitaka Yamamoto¹

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.

Developmental Biology

An Efficient Method to Obtain Dedifferentiated Fat Cells

Hiroaki Taniguchi^*1, Tomohiko Kazama^*1, Kazuhiro Hagikura^*1, Chii Yamamoto¹, Minako Kazama¹, Yuki Nagaoka¹, Taro Matsumoto¹

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.

Developmental Biology

Imaging of Cell Shape Alteration and Cell Movement in Drosophila Gastrulation Using DE-cadherin Reporter Transgenic Flies

M. Rezaul Karim¹, Tomohiro Haruta², Taro Matsumoto³, Hiroki Oda², 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.

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 Matsumoto³, Hidesato Ogawa¹, 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.

Biochemistry

Manipulating Living Cells to Construct Stable 3D Cellular Assembly Without Artificial Scaffold

Takehiro Yamazaki^*1, Hiroaki Taniguchi^*2, Shoto Tsuji¹, Shiho Sato¹, Takahiro Kenmotsu¹, Kenichi Yoshikawa¹, Koichiro Sadakane¹

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.

Developmental Biology

Neurogenesis Using P19 Embryonal Carcinoma Cells

Paweł Leszczyński¹, Magdalena Śmiech¹, Aamir S. Teeli¹, Aleksandra Zołocińska², Anna Słysz², Zygmunt Pojda², Mariusz Pierzchała³, Hiroaki Taniguchi¹

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.

Medicine

Novel Methods for Intranasal Administration Under Inhalation Anesthesia to Evaluate Nose-to-Brain Drug Delivery

Takanori Kanazawa^*1, Mitsuyoshi Fukuda^*1, Naoto Suzuki¹, Toyofumi Suzuki¹

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.

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.