Juntendo University View Institution's Website 7 articles published in JoVE Immunology and Infection Adjuvant Activity of Mycobacterium paratuberculosis in Enhancing the Immunogenicity of Autoantigens During Experimental Autoimmune Encephalomyelitis Davide Cossu1,2, Yuji Tomizawa1, Eiichi Momotani1, Kazumasa Yokoyama1, Nobutaka Hattori1,3 1Department of Neurology, Juntendo University, 2Biomedical Research Core Facilities, Juntendo University, 3Neurodegenerative Disorders Collaborative laboratory, RIKEN Center for Brain Science Here, we present an alternative protocol to actively induce experimental autoimmune encephalomyelitis in C57BL/6 mice, using the immunogenic epitope myelin oligodendrocyte glycoprotein (MOG)35-55 suspended in incomplete Freund's adjuvant containing the heat-killed Mycobacterium avium subspecies paratuberculosis. Biology Application of Passive Head Motion to Generate Defined Accelerations at the Heads of Rodents Takahiro Maekawa*1, Naoyoshi Sakitani*1, Youngjae Ryu1, Atsushi Takashima2, Shuhei Murase1, Julius Fink3, Motoshi Nagao1, Toru Ogata4, Masahiro Shinohara1, Yasuhiro Sawada5 1Department of Rehabilitation for Motor Functions, National Rehabilitation Center for Persons with Disabilities, 2Department of Assistive Technology, National Rehabilitation Center for Persons with Disabilities, 3Department of Metabolism and Endocrinology, Graduate School of Medicine, Juntendo University, 4Department of Rehabilitation Medicine, Graduate School of Medicine, The University of Tokyo, 5Department of Clinical Research, National Rehabilitation Center for Persons with Disabilities The present protocol describes a custom-designed ''passive head motion'' system, which reproduces mechanical accelerations at rodents' heads generated during their treadmill running at moderate velocities. It allows dissecting mechanical factors/elements from the beneficial effects of physical exercise. Neuroscience A Tissue Clearing Method for Neuronal Imaging from Mesoscopic to Microscopic Scales Kenta Yamauchi1,2, Shinichiro Okamoto1,2,3, Megumu Takahashi1,2,4,5, Masato Koike2,3, Takahiro Furuta6, Hiroyuki Hioki1,2,7 1Department of Neuroanatomy, Juntendo University Graduate School of Medicine, 2Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, 3Advanced Research Institute for Health Sciences, Juntendo University, 4Department of Neuroscience, Graduate School of Medicine, Kyoto University, 5Japan Society for the Promotion of Science, 6Department of Oral Anatomy and Neurobiology, Graduate School of Dentistry, Osaka University, 7Department of Multi-Scale Brain Structure Imaging, Juntendo University Graduate School of Medicine The protocol provides a detailed method of neuronal imaging in brain slice using a tissue clearing method, ScaleSF. The protocol includes brain tissue preparation, tissue clarification, handling of cleared slices and confocal laser scanning microscopy imaging of neuronal structures from mesoscopic to microscopic levels. Neuroscience Cheek Injection Model for Simultaneous Measurement of Pain and Itch-related Behaviors Yu Yamanoi1,2,3,4, Hiroki Kittaka1,2,5, Makoto Tominaga1,2,3,6 1Thermal Biology Group, Exploratory Research Center on Life and Living Systems (ExCELLS), 2Division of Cell Signaling, National Institute for Physiological Sciences, 3Department of Physiological Sciences, Graduate University for Advanced Studies (SOKENDAI), 4Research Laboratory, Ikedamohando Co., Ltd., 5Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, 6Institute for Environmental and Gender-Specific Medicine, Juntendo University Typically, the mouse neck injection model is used for evaluate pruritogen-induced scratch behaviors. However, the model provides information only on itch, not pain. Here, a cheek injection model is introduced in mice which can be used to simultaneously measure pain and itch-related behaviors. Medicine Lung Fixation under Constant Pressure for Evaluation of Emphysema in Mice Keiko Karasutani1,2, Hario Baskoro1, Tadashi Sato1, Naoko Arano1, Yohei Suzuki1, Aki Mitsui1, Naoko Shimada1, Yuzo Kodama1, Kuniaki Seyama1, Yoshinosuke Fukuchi1, Kazuhisa Takahashi1 1Department of Respiratory Medicine, Juntendo University Graduate School of Medicine, Juntendo University, 2Pharmaceutical Planning Group, Otsuka Pharmaceutical Co., Ltd. Presented here is a useful protocol for lung fixation that creates a stable condition for histological evaluate of lung specimens from a mouse model of emphysema. The main advantage of this model is that it can fix many lungs with the same constant pressure without lung collapse or deflation. Cancer Research High-sensitivity Detection of Micrometastases Generated by GFP Lentivirus-transduced Organoids Cultured from a Patient-derived Colon Tumor Yu Okazawa1,2, Kosuke Mizukoshi1,2, Yu Koyama2,4, Shoki Okubo5, Hiromitsu Komiyama1, Yutaka Kojima1, Michitoshi Goto1, Sonoko Habu3, Okio Hino2, Kazuhiro Sakamoto1, Akira Orimo2 1Department of Coloproctological Surgery, Juntendo University Faculty of Medicine, 2Department of Molecular Pathogenesis, Juntendo University Faculty of Medicine, 3Atopy Research Center, Juntendo University Faculty of Medicine, 4Department of Oral Pathobiological Science and Surgery, Tokyo Dental College, 5Department of Gastroenterology, Juntendo University Faculty of Medicine To allow highly sensitive detection of the disseminating human colorectal cancer (CRC) cells colonizing tissues, we herein show a protocol for efficient transduction of green fluorescent protein (GFP) lentiviral particles into PDX-derived CRC organoid cells prior to their injection into recipient mice, with stereo-fluorescence microscopic observation. Medicine Experimental Generation of Carcinoma-Associated Fibroblasts (CAFs) from Human Mammary Fibroblasts Urszula M. Polanska*1, Ahmet Acar*1, Akira Orimo1,2 1CR-UK Stromal-Tumour Interaction Group, Paterson Institute for Cancer Research, University of Manchester, 2Atopy Research Center, Juntendo University Carcinoma-associated fibroblasts (CAFs) rich in myofibroblasts present within the tumour stroma, play a major role in driving tumour progression. We developed a coimplantation tumour xengraft model for experimentally generating CAFs from human mammary fibroblasts. The protocol describes how to establish CAF myofibroblasts that acquire an ability to promote tumourigenesis.