Jichi Medical University 12 articles published in JoVE Medicine Delivery of Exogenous Artificially Synthesized miRNA Mimic to the Kidney Using Polyethylenimine Nanoparticles in Several Kidney Disease Mouse Models Katsunori Yanai*1, Shohei Kaneko1, Hiroki Ishii1, Akinori Aomatsu1, Yoshiyuki Morishita*1 1Division of Nephrology, First Department of Integrated Medicine, Saitama Medical Center, Jichi Medical University Here, we deliver exogenous artificially synthesized miRNA mimics to the kidney via tail vein injection of a nonviral vector and polyethylenimine nanoparticles in several kidney disease mouse models. This led to significant overexpression of target miRNA in the kidney, resulting in inhibited progression of kidney disease in several mouse models. Genetics Quantitative Real-Time Polymerase Chain Reaction Evaluation of MicroRNA Expression in Kidney and Serum of Mice with Age-Dependent Renal Impairment Katsunori Yanai1, Shohei Kaneko1, Hiroki Ishii1, Akinori Aomatsu1,2, Kenichi Ishibashi3, Yoshiyuki Morishita1 1Division of Nephrology, First Department of Integrated Medicine, Saitama Medical Center, Jichi Medical University, 2Division of Intensive Care Unit, First Department of Integrated Medicine, Saitama Medical Center, Jichi Medical University, 3Department of Medical Physiology, Meiji Pharmaceutical University We present a method for evaluating microRNA expression in the kidney and serum of mice with age-dependent renal impairment by quantitative reverse-transcription polymerase chain reaction. Medicine Sarcomere Shortening of Pluripotent Stem Cell-Derived Cardiomyocytes using Fluorescent-Tagged Sarcomere Proteins. Razan E. Ahmed*1, Nawin Chanthra*1, Tatsuya Anzai1,2, Keiichiro Koiwai3,4, Tomoki Murakami4, Hiroaki Suzuki4, Yutaka Hanazono1, Hideki Uosaki1 1Division of Regenerative Medicine, Center for Molecular Medicine, Jichi Medical University, 2Department of Pediatrics, Jichi Medical University, 3Institute of Global Innovation Research, Tokyo University of Agriculture and Technology, 4Department of Precision Mechanics, Faculty of Science and Engineering, Chuo University This method can be used to examine sarcomere shortening using pluripotent stem cell-derived cardiomyocytes with fluorescent-tagged sarcomere proteins. Medicine Measurement of Tissue Oxygenation Using Near-Infrared Spectroscopy in Patients Undergoing Hemodialysis Kiyonori Ito*1, Susumu Ookawara*1, Takayuki Uchida2, Hideyuki Hayasaka2, Masaya Kofuji2, Haruhisa Miyazawa1, Akinori Aomatsu1, Yuichiro Ueda1, Keiji Hirai1, Yoshiyuki Morishita1 1Division of Nephrology, First Department of Integrated Medicine, Saitama Medical Center, Jichi Medical University, 2Department of clinical engineering, Saitama Medical Center, Jichi Medical University We present a protocol to measure regional oxygen saturation (rSO2) in hemodialysis (HD) patients by using a near-infrared spectroscopy monitor. The rSO2 value is an index of tissue oxygenation. This noninvasive and real-time monitoring could be useful for confirming changes in organ oxygenation during HD. Genetics Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ureteral Obstruction Katsunori Yanai1, Shohei Kaneko1, Hiroki Ishii1, Akinori Aomatsu1,2, Kiyonori Ito1, Keiji Hirai1, Susumu Ookawara1, Kenichi Ishibashi3, Yoshiyuki Morishita1 1Division of Nephrology, First Department of Integrated Medicine, Saitama Medical Center, Jichi Medical University, 2Division of Intensive Care Unit, First Department of Integrated Medicine, Saitama Medical Center, Jichi Medical University, 3Department of Medical Physiology, Meiji Pharmaceutical University We describe a method for evaluating the microRNA expression in the kidneys of mice with unilateral ureteral obstruction (UUO) by quantitative reverse-transcription polymerase chain reaction. This protocol is suitable for studying kidney microRNA expression profiles in mice with UUO and in the context of other pathological conditions. Immunology and Infection Detection of MicroRNA Expression in the Kidneys of Immunoglobulin A Nephropathic Mice Shohei Kaneko1, Katsunori Yanai1, Hiroki Ishii1, Akinori Aomatsu1,2, Kiyonori Ito1, Keiji Hirai1, Susumu Ookawara1, Kenichi Ishibashi3, Yoshiyuki Morishita1 1Division of Nephrology, First Department of Integrated Medicine, Saitama Medical Center, Jichi Medical University, 2Division of Intensive Care Unit, First Department of Integrated Medicine, Saitama Medical Center, Jichi Medical University, 3Department of Medical Physiology, Meiji Pharmaceutical University microRNAs are involved in the pathogenesis of IgA nephropathy. We have developed a reliable method for detecting microRNA expression levels in the kidneys of an IgA nephropathy mouse model (HIGA mice). This new method will facilitate to check for miRNAs involvement in IgA nephropathy. Genetics Use of Freeze-thawed Embryos for High-efficiency Production of Genetically Modified Mice Hirofumi Nishizono*1,2,3, Mohamed Darwish*4,5, Hideki Uosaki6,7, Nanami Masuyama8,9,10, Motoaki Seki8,11, Hiroyuki Abe3, Nozomu Yachie8,9,10,12,13, Ryohei Yasuda1 1Max Planck Florida Institute for Neuroscience, 2Life Science Research Center, University of Toyama, 3Department of Biochemical Engineering, Graduate School of Science and Engineering, Yamagata University, 4Graduate School of Innovative Life Science, University of Toyama, 5Department of Biochemistry, Faculty of Pharmacy, Cairo University, 6Division of Regenerative Medicine, Center for Molecular Medicine, Jichi Medical University, 7Division of Stem Cell Research and Drug Development, Center for Development of Advanced Medical Technology, Jichi Medical University, 8Synthetic Biology Division, Research Center for Advanced Science and Technology, University of Tokyo, 9Institute for Advanced Biosciences, Keio University, 10Graduate School of Media and Governance, Keio University, 11Department of Molecular Oncology, Graduate School of Medicine, Chiba University, 12Department of Biological Sciences, School of Science, University of Tokyo, 13College of Arts and Sciences, University of Tokyo Here, we present a modified method for cryopreservation of one-cell embryos as well as a protocol that couples the use of freeze-thawed embryos and electroporation for the efficient generation of genetically modified mice. Behavior Low-cost Protocol of Footprint Analysis and Hanging Box Test for Mice Applied the Chronic Restraint Stress Hiroki Sugimoto1, Kiyoshi Kawakami1 1Division of Biology, Center for Molecular Medicine, Jichi Medical University The low-cost protocol consisting of footprint analysis and hanging box test after restraint stress is useful for evaluating the movement disorders of mouse model. Cancer Research Neutrophil Extracellular Traps Generated by Low Density Neutrophils Obtained from Peritoneal Lavage Fluid Mediate Tumor Cell Growth and Attachment Rihito Kanamaru1, Hideyuki Ohzawa1, Hideyo Miyato1, Hironori Yamaguchi1, Yoshinori Hosoya1, Alan Kawarai Lefor1, Naohiro Sata1, Joji Kitayama1 1Department of Gastrointestinal Surgery, Jichi Medical University Here, we present a method in which human low-density neutrophils (LDN), recovered from postoperative peritoneal lavage fluid, produce massive neutrophil extracellular traps (NETs) and efficiently trap free tumor cells that subsequently grow. Biology Partial Bile Duct Ligation in the Mouse: A Controlled Model of Localized Obstructive Cholestasis Shinichiro Yokota1,2, Yoshihiro Ono2, Toshimasa Nakao2, Peng Zhang3, George K. Michalopoulos4,5, Zahida Khan3,4,5,6 1Department of Surgery, Jichi Medical University, 2Department of Surgery, University of Pittsburgh School of Medicine, 3Department of Pediatrics, University of Pittsburgh School of Medicine, 4Department of Pathology, University of Pittsburgh School of Medicine, 5Pittsburgh Liver Research Center, University of Pittsburgh, 6McGowan Institute for Regenerative Medicine, University of Pittsburgh Here, we present partial bile duct ligation as a surgical model of liver injury and regeneration in rodents. Genetics Detection of microRNA Expression in Peritoneal Membrane of Rats Using Quantitative Real-time PCR Keiji Hirai1, Hiromichi Yoshizawa2, Toshimi Imai2, Yusuke Igarashi2, Ichiro Hirahara2, Susumu Ookawara2, Kenichi Ishibashi3, Yoshiyuki Morishita1 1Division of Nephrology, First Department of Integrated Medicine, Saitama Medical Center, Jichi Medical University, 2Division of Nephrology, Department of Internal Medicine, Jichi Medical University, 3Department of Medical Physiology, Meiji Pharmaceutical University Here we present a protocol for the detection of microRNA expression in rat peritoneal membrane using quantitative real-time reverse-transcription polymerase chain reaction. This method is suitable for studying the microRNA expression profile in rat peritoneal membrane in several pathological conditions. Biology Preparation of Rat Serum Suitable for Mammalian Whole Embryo Culture Masanori Takahashi1,2, Sayaka Makino3, Takako Kikkawa3, Noriko Osumi3 1Graduate School of Medicine, Jichi Medical University, 2Division of Biology, Center for Molecular Medicine, Jichi Medical University, 3Department of Developmental Neuroscience, United Center for Advanced Research and Translational Medicine (ART), Tohoku University School of Medicine Mammalian whole embryo culture (WEC) is widely used in teratology and developmental biology. Immediately centrifuged rat serum is commonly provided as a medium for both mouse and rat WEC. In this video, we demonstrate our standard protocol for the preparation of high-quality rat serum suitable for mammalian WEC.
