Osaka Medical and Pharmaceutical University

3 ARTICLES PUBLISHED IN JoVE

Biochemistry

Nucleoside Triphosphate Hydrolases Assay in Toxoplasm gondii and Neospora caninum for High-Throughput Screening using a Robot Arm

Riho Kurata^*1, Masamitsu Harada^*2, Jun Nagai^*3, Xiaofeng Cui^*4, Takayuki Isagawa⁵, Hiroaki Semba⁶, Yasuhiro Yoshida⁷, Norihiko Takeda⁸, Koji Maemura⁹, Tomo Yonezawa³

¹Education and Research Center for Pharmaceutical Sciences, Faculty of Pharmacy, Osaka Medical and Pharmaceutical University, ²Independent Scholar, Tokyo, Japan, ³Division of Functional Genomics and Therapeutic Innovation, Research Center for Advanced Genomics, Graduate School of Biomedical Sciences, Nagasaki University, ⁴School of Chemistry, Chemical Engineering and Life Sciences, School of Materials and Engineering, Wuhan University of Technology, ⁵Data Science Center, Jichi Medical University, ⁶Department of Cardiovascular Medicine, The Cardiovascular Institute, ⁷Department of Immunology and Parasitology, University of Occupational and Environmental Health, ⁸Division of Cardiology and Metabolism, Center for Molecular Medicine, Jichi Medical University, ⁹Department of Cardiovascular Medicine, Graduate School of Biomedical Sciences, Nagasaki University Hospital

Toxoplasma gondii and Neospora caninum infections are found in humans and animals and lead to serious health issues. The two parasites share similar nucleoside triphosphate hydrolases and play important roles in propagation and survival. We established a high-standard assay of the enzymes requiring robot arm usage.

Biology

The 3D Culturing of Organoids from Murine Intestinal Crypts and a Single Stem Cell for Organoid Research

Yuta Takase¹, Kazuto Fujishima^2,3, Toshio Takahashi¹

¹Suntory Foundation for Life Sciences, Bioorganic Research Institute, ²Institute for Integrated Cell-Material Sciences (KUIAS-iCeMS), Kyoto University, ³Faculty of Medicine, Osaka Medical and Pharmaceutical University

We describe a protocol to isolate murine small intestinal crypts and culture intestinal 3D organoids from the crypts. Additionally, we describe a method to generate organoids from a single intestinal stem cell in the absence of a sub-epithelial cellular niche.

Neuroscience

Live Imaging of Synaptic Vesicle Recycling in the Neuromuscular Junction of Dissected Larval Zebrafish

Yoshihiro Egashira¹, Fumihito Ono¹

¹Department of Physiology, Osaka Medical and Pharmaceutical University

The zebrafish larval neuromuscular junction is an attractive model for studying synaptic physiology. It is amenable to many experimental techniques, including electrophysiology and optical imaging. Here, we describe a protocol for imaging synaptic transmission using a pHluorin-based probe under an upright epifluorescence microscope.