University of Yamanashi 7 articles published in JoVE Neuroscience In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window Satoshi Manita1, Eiji Shigetomi2,3, Haruhiko Bito4, Schuichi Koizumi2,3, Kazuo Kitamura1 1Department of Neurophysiology, Faculty of Medicine, University of Yamanashi, 2Department of Neuropharmacology, Faculty of Medicine, University of Yamanashi, 3Yamanashi GLIA center, Interdisciplinary Graduate School of Medicine, University of Yamanashi, 4Department of Neurochemistry, Graduate School of Medicine, The University of Tokyo The present protocol describes making a large (6 x 3 mm2) cranial window using food wrap, transparent silicone, and cover glass. This cranial window allows in vivo wide-field and two-photon calcium imaging experiments in the same mouse. Neuroscience Transplantation of Human Induced Pluripotent Stem Cell-Derived Microglia in Immunocompetent Mice Brain via Non-Invasive Transnasal Route Bijay Parajuli1,2, Youichi Shinozaki1,2, Eiji Shigetomi1,2, Schuichi Koizumi1,2 1Department of Neuropharmacology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, 2GLIA Center, University of Yamanashi The protocol presented here allows the transplantation of induced pluripotent stem cell-derived human microglia (iPSMG) into the brain via a transnasal route in immunocompetent mice. The method for the preparation and transnasal transplantation of cells and the administration of cytokine mixture for the maintenance of iPSMG is shown. Biochemistry Isolation and Analysis of Plasma Lipoproteins by Ultracentrifugation Manabu Niimi1, Haizhao Yan1, Yajie Chen1, Yao Wang2, Jianglin Fan1,2 1Department of Molecular Pathology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, 2School of Biotechnology and Health Sciences, Wuyi University Several methods have been used for analyzing plasma lipoproteins; however, ultracentrifugation is still one of the most popular and reliable methods. Here, we describe a method regarding how to isolate lipoproteins from plasma using sequential density ultracentrifugation and how to analyze the apolipoproteins for both diagnostic and research purposes. Chemistry Preparation of Polyoxometalate-based Photo-responsive Membranes for the Photo-activation of Manganese Oxide Catalysts Akira Yamaguchi1,5, Toshihiro Takashima2, Kazuhito Hashimoto1,6, Ryuhei Nakamura3,4 1Department of Applied Chemistry, The University of Tokyo, 2Clean Energy Research Center, University of Yamanashi, 3Biofunctional Catalyst Research Team, RIKEN Center for Sustainable Resource Science, 4Earth-Life Science Institute (ELSI), Tokyo Institute of Technology, 5Department of Materials Science and Engineering, Tokyo Institute of Technology, 6National Institute for Materials Science Here, we present a protocol to prepare charge transfer chromophores based on a polyoxometalate/polymer composite membrane. Developmental Biology Zygotic Fluorescence Recovery After Photo-bleaching Analysis for Chromatin Looseness That Allows Full-term Development Masatoshi Ooga1,3, Satoshi Funaya2, Fugaku Aoki2, Teruhiko Wakayama1,3 1Faculty of Life and Environmental Sciences, Department of Biotechnology, University of Yamanashi, 2Department of Integrated Bioscience, Graduate School of Frontier Sciences, University of Tokyo, 3Advanced Biotechnology Center, University of Yamanashi Chromatin looseness appears to be involved in the developmental potential of blastomeres. However, it is not known whether chromatin looseness can be used as a reliable index for the developmental potential for embryos. Here, an experimental system in which chromatin looseness-evaluated zygotes can develop to full term has been described. Neuroscience Observation of the Ciliary Movement of Choroid Plexus Epithelial Cells Ex Vivo Takafumi Inoue1, Keishi Narita2, Yuta Nonami1, Hideki Nakamura1, Sen Takeda2 1Department of Life Science and Medical Bioscience, Faculty of Science and Engineering, Waseda University, 2Department of Anatomy and Cell Biology, Interdisciplinary Graduate School of Medicine & Engineering, University of Yamanashi In this study, a detailed light microscopic technique was optimized for real-time observation and analysis of the motion of CPEC cilia ex vivo together with an electron microscopic method for ultrastructural analysis. Medicine Production of Apolipoprotein C-III Knockout Rabbits using Zinc Finger Nucleases Dongshan Yang1, Jifeng Zhang1, Jie Xu1, Tianqing Zhu1, Yanbo Fan1, Jianglin Fan2, Y. Eugene Chen1 1Center for Advanced Models for Translational Sciences and Therapeutics, Department of Internal Medicine, University of Michigan Medical Center, 2Department of Molecular Pathology, University of Yamanashi Recent development in gene targeting tools makes production of knockout (KO) rabbits possible. In the present work, we generated five Apolipoprotein (Apo) C-III KO rabbits using Zinc Finger Nucleases (ZFN). This work demonstrated that ZFN is a highly efficient method to produce KO rabbits.
