Graduate University for Advanced Studies (SOKENDAI)

4 ARTICLES PUBLISHED IN JoVE

Neuroscience

Ablation of a Neuronal Population Using a Two-photon Laser and Its Assessment Using Calcium Imaging and Behavioral Recording in Zebrafish Larvae

Akira Muto¹, Koichi Kawakami¹

¹Division of Molecular and Developmental Biology, National Institute of Genetics, Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies)

Here, we present a protocol to ablate a genetically labeled subpopulation of neurons by a two-photon laser from Zebrafish larvae.

Neuroscience

Cheek Injection Model for Simultaneous Measurement of Pain and Itch-related Behaviors

Yu Yamanoi^1,2,3,4, Hiroki Kittaka^1,2,5, Makoto Tominaga^1,2,3,6

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.

Developmental Biology

Isolation of Murine Spermatogenic Cells using a Violet-Excited Cell-Permeable DNA Binding Dye

Yu-Han Yeh^*1,2, Mengwen Hu^*1,2, Toshinori Nakagawa^3,4, Akihiko Sakashita^1,2, Shosei Yoshida^3,4, So Maezawa^5,6, Satoshi H. Namekawa^1,2

¹Division of Reproductive Sciences, Division of Developmental Biology, Perinatal Institute, Cincinnati Children's Hospital Medical Center, ²Department of Pediatrics, University of Cincinnati College of Medicine, ³Division of Germ Cell Biology, National Institute for Basic Biology, National Institutes of Natural Sciences, ⁴Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies (Sokendai), ⁵Department of Animal Science and Biotechnology, School of Veterinary Medicine, Azabu University, ⁶Faculty of Science and Technology, Department of Applied Biological Science, Tokyo University of Science

Here we present a simple and efficient method to isolate live meiotic and post-meiotic germ cells from adult mouse testes. Using a low-cytotoxicity, violet-excited DNA binding dye and fluorescence-activated cell sorting, one can isolate highly enriched spermatogenic cell populations for many downstream applications.

Neuroscience

Optogenetic Phase Transition of TDP-43 in Spinal Motor Neurons of Zebrafish Larvae

Kazuhide Asakawa¹, Hiroshi Handa¹, Koichi Kawakami^2,3

¹Department of Chemical Biology, Tokyo Medical University, ²Division of Molecular and Developmental Biology, National Institute of Genetics, ³Department of Genetics, Graduate University for Advanced Studies (SOKENDAI)

We describe a protocol to induce phase transition of TAR DNA-binding protein 43 (TDP-43) by light in the spinal motor neurons using zebrafish as a model.