Japan Science and Technology Agency (JST)

4 ARTICLES PUBLISHED IN JoVE

Medicine

Visualization and Analysis of Blood Flow and Oxygen Consumption in Hepatic Microcirculation: Application to an Acute Hepatitis Model

Kosuke Tsukada¹, Makoto Suematsu^2,3

¹Department of Applied Physics and Physico-Informatics, Keio University, ²Department of Biochemistry, School of Medicine, Keio University, ³Exploratory Research for Advanced Technology (ERATO), Suematsu Gas Biology Project, Japan Science and Technology Agency (JST)

An optical system was developed to visualize hepatic microcirculation with FITC-labeled erythrocytes and to measure the partial pressure of oxygen in the microvessels with laser-assisted phosphorimetry. This method can be used to investigate physiological and pathological mechanisms by analyzing microvascular structure, diameter, blood flow velocity, and oxygen tension.

Neuroscience

Quadruple Immunostaining of the Olfactory Bulb for Visualization of Olfactory Sensory Axon Molecular Identity Codes

Naoki Ihara¹, Yuji Ikegaya^1,2, Haruki Takeuchi^1,3

¹Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, ²Center for Information and Neural Networks, National Institute of Information and Communications Technology, ³PRESTO, Japan Science and Technology Agency (JST)

Olfactory sensory neurons express a wide variety of axon-sorting molecules to establish proper neural circuitry. This protocol describes an immunohistochemical staining method to visualize combinatorial expressions of axon-sorting molecules at the axon termini of olfactory sensory neurons.

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 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.

JoVE Journal

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

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.