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Tokyo University of agriculture and technology

10 ARTICLES PUBLISHED IN JoVE

Neuroscience

Assaying Surface Expression of Chemosensory Receptors in Heterologous Cells

Sandeepa Dey¹, Senmiao Zhan², Hiroaki Matsunami¹

¹Department of Molecular Genetics and Molecular Biology , Duke University, ²Department of Chemistry, Duke University

Here we demonstrate a protocol to carry out live cell staining that can be used to detect odorant receptors on the surface of HEK293T cells conveniently. In addition, it may also be used to assay for surface expression of other chemosensory receptors or GPCRs.

Behavior

A Mouse Model of Subchronic and Mild Social Defeat Stress for Understanding Stress-induced Behavioral and Physiological Deficits

Tatsuhiko Goto^1,2, Atsushi Toyoda^1,2,3

¹College of Agriculture, Ibaraki University, ²Cooperation between Agriculture and Medical Science, Ibaraki University, ³United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology

Here, methods for developing a mouse model of subchronic and mild social defeat stress are described and used to investigate the pathogenic features of depression including hyperphagia- and polydipsia-like symptoms following increased body weight.

Behavior

Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test

Akari Asaba¹, Masahiro Kato³, Nobuyoshi Koshida², Takefumi Kikusui¹

¹School of Veterinary Medicine, Azabu University, ²Division of Electric and Information Engineering, Tokyo University of Agriculture and Technology, ³Kato Acoustics Consulting Office

Ultrasonic vocalizations (USVs) in mice differ depending on age, sex, condition, and genetic background. Using two ultrasound emitters broadcasting simultaneously in different locations, this two-choice test can evaluate murine recognition and preference responses to different characteristics of USVs.

Chemistry

Construction of Models for Nondestructive Prediction of Ingredient Contents in Blueberries by Near-infrared Spectroscopy Based on HPLC Measurements

Wenming Bai¹, Norio Yoshimura¹, Masao Takayanagi¹, Jingai Che², Naomi Horiuchi², Isao Ogiwara³

¹United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, ²Faculty of Agriculture, Tokyo University of Agriculture and Technology, ³Institute of Agriculture, Tokyo University of Agriculture and Technology

We present here a protocol to construct and validate models for nondestructive prediction of total sugar, total organic acid, and total anthocyanin content in individual blueberries by near-infrared spectroscopy.

Neuroscience

Live-cell Measurement of Odorant Receptor Activation Using a Real-time cAMP Assay

Yuetian Zhang^*1, Yi Pan^*1, Hiroaki Matsunami^2,3, Hanyi Zhuang^1,4

¹Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education, Shanghai Jiao Tong University School of Medicine, ²Department of Molecular Genetics and Microbiology, Duke University Medical Center, ³Department of Neurobiology, Duke Institute for Brain Sciences, Duke University Medical Center, ⁴Institute of Health Science, Chinese Academy of Science/Shanghai Jiao Tong University School of Medicine

Characterizing the function of odorant receptors serves an indispensable part in the deorphanization process. We describe a method to measure the activation of odorant receptors in real time using a cAMP assay.

Environment

Tracking Infiltration Front Depth Using Time-lapse Multi-offset Gathers Collected with Array Antenna Ground Penetrating Radar

Hirotaka Saito¹, Seiichiro Kuroda², Toshiki Iwasaki¹, Haruyuki Fujimaki³, Nobuhito Nagai⁴, Jacopo Sala⁵

¹Graduate School of Agriculture, Tokyo University of Agriculture and Technology, ²National Institute for Rural Engineering, National Agriculture and Food Research Organization, ³Arid Land Research Center, Tottori University, ⁴GeoFive Co. Ltd, ⁵3D-Radar

Here we present a Ground Penetrating Radar (GPR) system based on a ground-coupled, densely populated antenna array for monitoring the dynamic process of subsurface water infiltration. A time-lapse radar image of the infiltration process allowed estimating the depth of the wetting front during the course of the infiltration process.

Neuroscience

Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase

Claire A. de March¹, Yosuke Fukutani^1,2, Aashutosh Vihani^1,3, Hitoshi Kida^1,4, Hiroaki Matsunami^1,3,5,6

¹Department of Molecular Genetics and Microbiology, Duke University Medical Center, ²Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, ³Department of Neurobiology, Duke University Medical Center, ⁴Department of Mechanical Systems, Engineering, Tokyo University of Agriculture and Technology, ⁵Institute of Global Innovation Research, Tokyo University of Agriculture and Technology, ⁶Duke Institute for Brain Sciences, Duke University

Physiologically, odorant receptors are activated by odorant molecules inhaled in the vapor phase. However, most in vitro systems utilize liquid phase odorant stimulation. Here, we present a method that allows real-time in vitro monitoring of odorant receptor activation upon odorant stimulation in vapor phase.

Medicine

Sarcomere Shortening of Pluripotent Stem Cell-Derived Cardiomyocytes using Fluorescent-Tagged Sarcomere Proteins.

Razan E. Ahmed^*1, Nawin Chanthra^*1, Tatsuya Anzai^1,2, Keiichiro Koiwai^3,4, 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.

Environment

Mass-Rearing and Molecular Studies in Tortricidae Pest Insects

Hiroshi Arai¹, Yuna Ishitsubo¹, Madoka Nakai¹, Maki N. Inoue¹

¹Department of Applied Biological Science, Tokyo University of Agriculture and Technology

The present protocol describes the rearing method of tortricid pest insects in the laboratories. The procedures to distinguish insects' sex and extract nucleic acids for high throughput sequencing are established using two tortricid pests.

Bioengineering

Cellular Affinity of Particle-Stabilized Emulsion to Boost Antigen Internalization

Fengqiang Cao^*1,2,3, Yali Ming^*3,4, Weixiang Gao^3,4, Jia Ge³, Kenji Ogino^1,2

¹Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, ²Institute of Global Innovation Research, Tokyo University of Agriculture and Technology, ³State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, ⁴University of Chinese Academy of Sciences

To rationally design efficient adjuvants, we developed poly-lactic-co-glycolic acid nanoparticle-stabilized Pickering emulsion (PNPE). The PNPE possessed unique softness and a hydrophobic interface for potent cellular contact and offered high-content antigen loading, improving the cellular affinity of the delivery system to antigen-presenting cells and inducing efficient internalization of antigens.