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Live-Cell Single-Molecule Imaging with Optogenetics Reveals Dynamics of a Neuronal Activity-Dependent Transcription Factor

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Single Molecule Microscopy in Neurobiology

Part of the book series: Neuromethods ((NM,volume 154))

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Abstract

Powerful imaging techniques have been developed to investigate the spatiotemporal dynamics of molecular players that are involved in various biological functions. A fine-tuned single-molecule imaging technique allows us to study the movement of transcription factors in the cell nucleus. Our technique combined with optogenetics enables us to reveal neuronal activity-dependent dynamics of transcription factors in living cortical neurons. Here, we describe the detailed experimental procedures to study the transcriptional activity with physiological stimulation in living CNS neurons.

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Acknowledgments

This research was supported by MEXT KAKENHI on Innovative Areas “Mesoscopic Neurocircuitry” (No. 23115102) to N.Y., “Dynamic Regulation of Brain Function by Scrap and Build System” (No. 16H06460) to N.Y., “Cross-talk between moving cells and microenvironment as a basis of emerging order in multicellular system” (No. 23111516) to N.S., and Grant Nos. 20200009 to N.S. and 20300110 and 2330018 to N.Y. We thank Dr. Ian smith for critical reading of the manuscript and Dr. Yoshiyuki Arai for providing the PTA plug-in. We also thank Dr. Masatoshi Morimatsu and Dr. Toshio Yanagida for setting up the single-molecule microscopy system.

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Authors and Affiliations

  1. Laboratory of Cellular and Molecular Neurobiology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan

    Hironobu Kitagawa, Noriyuki Sugo & Nobuhiko Yamamoto

Authors
  1. Hironobu Kitagawa
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  2. Noriyuki Sugo
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  3. Nobuhiko Yamamoto
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Corresponding author

Correspondence to Nobuhiko Yamamoto .

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Editors and Affiliations

  1. Laboratory of Cellular and Molecular Neurobiology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan

    Nobuhiko Yamamoto

  2. Laboratory for Cell Polarity Regulation, RIKEN Center for Biosystems Dynamics Research, Suita, Osaka, Japan

    Yasushi Okada

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Kitagawa, H., Sugo, N., Yamamoto, N. (2020). Live-Cell Single-Molecule Imaging with Optogenetics Reveals Dynamics of a Neuronal Activity-Dependent Transcription Factor. In: Yamamoto, N., Okada, Y. (eds) Single Molecule Microscopy in Neurobiology . Neuromethods, vol 154. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0532-5_4

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  • DOI: https://doi.org/10.1007/978-1-0716-0532-5_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0531-8

  • Online ISBN: 978-1-0716-0532-5

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