NDLI: Rational design of a high-affinity, fast, red calcium indicator R-CaMP2.

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Rational design of a high-affinity, fast, red calcium indicator R-CaMP2.

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Rational design of a high-affinity, fast, red calcium indicator R-CaMP2.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Inoue, Masatoshi Takeuchi, Atsuya Horigane, Shin-ichiro Ohkura, Masamichi Gengyo-Ando, Keiko Fujii, Hajime Kamijo, Satoshi Takemoto-Kimura, Sayaka Kano, Masanobu Nakai, Junichi Kitamura, Kazuo Bito, Haruhiko
Description	Country affiliation: Japan Author Affiliation: Inoue M ( 1] Department of Neurochemistry, Graduate School of Medicine, University of Tokyo, Tokyo, Japan. [2] Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Tokyo, Japan.); Takeuchi A ( Department of Neurophysiology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.); Horigane S ( Department of Neurochemistry, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.); Ohkura M ( Brain Science Institute, Saitama University, Saitama, Japan.); Gengyo-Ando K ( Brain Science Institute, Saitama University, Saitama, Japan.); Fujii H ( Department of Neurochemistry, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.); Kamijo S ( 1] Department of Neurochemistry, Graduate School of Medicine, University of Tokyo, Tokyo, Japan. [2] Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Tokyo, Japan.); Takemoto-Kimura S ( 1] Department of Neurochemistry, Graduate School of Medicine, University of Tokyo, Tokyo, Japan. [2] Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency, Saitama, Japan.); Kano M ( Department of Neurophysiology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.); Nakai J ( Brain Science Institute, Saitama University, Saitama, Japan.); Kitamura K ( 1] Department of Neurophysiology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan. [2] Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency, Saitama, Japan.); Bito H ( 1] Department of Neurochemistry, Graduate School of Medicine, University of Tokyo, Tokyo, Japan. [2] Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Tokyo, Japan.)
Abstract	Fluorescent Ca(2+) reporters are widely used as readouts of neuronal activities. Here we designed R-CaMP2, a high-affinity red genetically encoded calcium indicator (GECI) with a Hill coefficient near 1. Use of the calmodulin-binding sequence of CaMKK- and CaMKK-ß in lieu of an M13 sequence resulted in threefold faster rise and decay times of Ca(2+) transients than R-CaMP1.07. These features allowed resolving single action potentials (APs) and recording fast AP trains up to 20-40 Hz in cortical slices. Somatic and synaptic activities of a cortical neuronal ensemble in vivo were imaged with similar efficacy as with previously reported sensitive green GECIs. Combining green and red GECIs, we successfully achieved dual-color monitoring of neuronal activities of distinct cell types, both in the mouse cortex and in freely moving Caenorhabditis elegans. Dual imaging using R-CaMP2 and green GECIs provides a powerful means to interrogate orthogonal and hierarchical neuronal ensembles in vivo.
File Format	HTM / HTML
ISSN	15487091
Issue Number	1
Volume Number	12
e-ISSN	15487105
Journal	Nature Methods
Language	English
Publisher	Nature Publishing Group
Publisher Date	2015-01-01
Publisher Place	United States
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Clinical Laboratory Techniques Calcium-calmodulin-dependent Protein Kinase Kinase Metabolism Indicators And Reagents Chemical Synthesis Action Potentials Physiology Animals Caenorhabditis Elegans Radiation Effects Calcium Calcium Signaling Calmodulin-binding Proteins Cells, Cultured Cerebral Cortex Cytology Fluorescent Dyes Hek293 Cells Hippocampus Humans Light Mice Neurons Patch-clamp Techniques Peptide Fragments Chemistry Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology Biotechnology

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