Saitama University View Institution's Website 6 articles published in JoVE Biology Wide-Field, Real-Time Imaging of Local and Systemic Wound Signals in Arabidopsis Takuya Uemura1, Jiaqi Wang1, Yuri Aratani1, Simon Gilroy2, Masatsugu Toyota1,2 1Department of Biochemistry and Molecular Biology, Saitama University, 2Department of Botany, University of Wisconsin Extracellular glutamate-triggered systemic calcium signaling is critical for the induction of plant defense responses to mechanical wounding and herbivore attack in plants. This article describes a method to visualize the spatial and temporal dynamics of both these factors using Arabidopsis thaliana plants expressing calcium- and glutamate-sensitive fluorescent biosensors. Biology Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography Kazuyoshi Murata1, Yasuko Kaneko2 1National Institute for Physiological Sciences, 2Graduate School of Science and Engineering, Saitama University This protocol describes how to visualize the transient DNA compaction in cyanobacteria. Synchronous cultivation, monitoring by fluorescence microscopy, rapid freezing, and high voltage cryo-electron tomography are used. A protocol for these methodologies is presented, and future applications and developments are discussed. Biology Real-time In Vivo Recording of Arabidopsis Calcium Signals During Insect Feeding Using a Fluorescent Biosensor Thomas R. Vincent1, James Canham1, Masatsugu Toyota2,3,4, Marieta Avramova1, Sam T. Mugford5, Simon Gilroy2, Anthony J. Miller1, Saskia Hogenhout5, Dale Sanders1 1Department of Metabolic Biology, John Innes Centre, Norwich Research Park, 2Department of Botany, University of Wisconsin, Madison, 3Department of Biochemistry and Molecular Biology, Saitama University, 4Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), 5Department of Cell and Developmental Biology, John Innes Centre, Norwich Research Park This protocol outlines a simple method for analyzing calcium signals in plants generated by feeding hemipteran insects, such as aphids. Arabidopsis thaliana transformed with the GFP calcium biosensor GCaMP3 allow for the real-time in vivo imaging of calcium dynamics with a high temporal and spatial resolution. Biology Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus Ke Ren1, Yidan Dai1, Kai Yi2, Masanobu Kinoshita1, Masahiro Itoh2, Ichiro Sakata3, Takafumi Sakai3, Shuang-Qin Yi1 1Department of Frontier Health Sciences, Graduate School of Human Health Sciences, Tokyo Metropolitan University, 2Department of Anatomy, Tokyo Medical University, 3Area of Regulatory Biology, Division of Life Science, Graduate School of Science and Engineering, Saitama University A whole-mount immunohistochemical approach, to visualize neurofilament protein expression in the extrahepatic biliary tract in Suncus murinus. is presented here. This protocol can be used to analyze the innervation of all visceral organs in S. murinus or other species. Neuroscience Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall Cagri Yalgin1,2, M. Rezaul Karim1,2, Adrian W. Moore1 1Disease Mechanism Research Core, RIKEN Brain Science Institute, 2Graduate School of Science and Engineering, Saitama University To understand how complex cell shapes, such as neuronal dendrites, are achieved during development, it is important to be able to accurately assay microtubule organization. Here we describe a robust immunohistological labeling method to examine microtubule organization of dendritic arborization neuron sensory dendrites, trachea, muscle, and other Drosophila larva body wall tissues. Neuroscience Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics M. Rezaul Karim1,2, Adrian W. Moore1 1Disease Mechanism Research Core, RIKEN Brain Science Institute, 2Graduate School of Science and Engineering, Saitama University The dendritic arborization sensory neurons of the Drosophila larval peripheral nervous system are useful models to elucidate both general and neuron class-specific mechanisms of neuron differentiation. We present a practical guide to generate and analyze dendritic arborization neuron genetic mosaics.