Saitama University

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 Toyota^{1,2 1}Department 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 Kaneko^{2 1}National 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 Toyota^2,3,4, Marieta Avramova¹, Sam T. Mugford⁵, Simon Gilroy², Anthony J. Miller¹, Saskia Hogenhout⁵, Dale Sanders^{1 1}Department 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 Yi^{1 1}Department 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 Yalgin^1,2, M. Rezaul Karim^1,2, Adrian W. Moore^{1 1}Disease 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 Karim^1,2, Adrian W. Moore^{1 1}Disease 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.