Doshisha University 10 articles published in JoVE Biochemistry Quantitative Microtubule Fractionation Technique to Separate Stable Microtubules, Labile Microtubules, and Free Tubulin in Mouse Tissues Ayaka Hagita-Tatsumoto1,2, Tomohiro Miyasaka1,2,3 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. Behavior Utilizing a Reconfigurable Maze System to Enhance the Reproducibility of Spatial Navigation Tests in Rodents Fumiya Sawatani1, Yuta Tamatsu1, Kaoru Ide1, Hirotsugu Azechi1, Susumu Takahashi1 1Laboratory of Cognitive and Behavioral Neuroscience, Graduate School of Brain Science, Doshisha University The present protocol describes a reconfigurable maze, a unique system for testing spatial navigation and behavioral phenotypes in rodents. The adaptability of this maze system enables the execution of various experiments in a single physical environment. The ease of structural rearrangement generates reliable and reproducible experimental results. Behavior A Task for Assessing the Impact of a Partner on the Speed and Accuracy of Motor Performance in Rats Yayoi Sekiguchi1,2, Toshimichi Hata3 1Graduate School of Psychology, Doshisha University, 2Japan Society for the Promotion of Science, 3Faculty of Psychology, Doshisha University A procedure to measure the speed and accuracy of rats’ motor performance in a social condition is described. The protocol enables us to investigate the effect of the mere presence of others on speed and accuracy of motor performance in one experiment. Neuroscience Visualization of Amyloid β Deposits in the Human Brain with Matrix-assisted Laser Desorption/Ionization Imaging Mass Spectrometry Masaya Ikegawa*1, Takashi Nirasawa*2, Nobuto Kakuda1, Tomohiro Miyasaka1, Yuki Kuzuhara1, Shigeo Murayama3, Yasuo Ihara4 1Department of Life and Medical Systems, Doshisha University, 2Bruker Daltonics K.K., 3The Brain Bank for Aging Research, Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology, 4Graduate School of Brain Science, Doshisha University Molecular imaging with matrix-assisted laser desorption/ionization-based imaging mass spectrometry (MALDI-IMS) allows the simultaneous mapping of multiple analytes in biological samples. Here, we present a protocol for the detection and visualization of amyloid β protein on brain tissues of Alzheimer's disease and cerebral amyloid angiopathy samples using MALDI-IMS. 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. Behavior Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage Akira Masuda1,2, Yuki Kobayashi1, Shigeyoshi Itohara1 1Laboratory for Behavioral Genetics, Center for Brain Science, RIKEN, 2Organization for Research Initiatives and Development, Doshisha University This paper describes a protocol for cognitive assessments for genetic models of the Alzheimer's disease using the IntelliCage system, which is a high throughput automated behavioral monitoring system with operant conditioning. Immunology and Infection Evaluation of Blood Lactate and Plasma Insulin During High-intensity Exercise by Antecubital Vein Catheterization Minas Nalbandian1, Zsolt Radak2, Masaki Takeda3 1Graduate School of medicine, Kyoto University, 2Research Institute of Sports Science, University of Physical Education, 3Graduate School of Sports and Health Science, Doshisha University Here we present a protocol to obtain several blood samples from the antecubital vein during high-intensity interval training. This protocol may be useful for the measurement of blood metabolites and endocrinal markers during exercise. 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 Taniguchi3,4,5 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. 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 Taniguchi3,4,5 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. 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.
