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Improving membrane voltage measurements using FRET with new fluorescent proteins

Nature Methods volume 5, pages 683–685 (2008)Cite this article

Abstract

We used two new coral fluorescent proteins as fluorescence resonance energy transfer (FRET) donor and acceptor to develop a voltage sensor, named Mermaid, that displays ∼40% changes in emission ratio per 100 mV, allowing for direct visualization of electrical activities in cultured excitable cells. Notably, Mermaid has fast on-off kinetics at warm (∼33 °C) temperatures and can report voltage spikes comparable to action potentials.

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Figure 1: Development and kinetics of the voltage sensor Mermaid.
Figure 2: Depolarization patterns in cultured excitable cells expressing Mermaid visualized in single sweeps.

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References

  1. Tsien, R.Y. Annu. Rev. Biochem. 67, 509–544 (1998).

    Article  CAS  Google Scholar 

  2. Miyawaki, A. Neuron 48, 189–199 (2005).

    Article  CAS  Google Scholar 

  3. Nagai, T. et al. Proc. Natl. Acad. Sci. USA 101, 10554–10559 (2004).

    Article  CAS  Google Scholar 

  4. Campbell, R.E. et al. Proc. Natl. Acad. Sci. USA 99, 7877–7882 (2002).

    Article  CAS  Google Scholar 

  5. Karasawa, S. et al. Biochem. J. 381, 307–312 (2004).

    Article  CAS  Google Scholar 

  6. Siegel, M.S. & Isacoff, E.Y. Neuron 4, 735–741 (1997).

    Article  Google Scholar 

  7. Guerrero, G. et al. Biophys. J. 83, 3607–3618 (2002).

    Article  CAS  Google Scholar 

  8. Sakai, R. et al. Eur. J. Neurosci. 12, 2314–2318 (2001).

    Article  Google Scholar 

  9. Ataka, K. et al. Biophys. J. 82, 509–516 (2002).

    Article  CAS  Google Scholar 

  10. Murata, Y. et al. Nature 435, 1239–1243 (2005).

    Article  CAS  Google Scholar 

  11. Dimitrov, D. et al. PLoS One 5, e440 (2007).

    Article  Google Scholar 

  12. Seoh, S.A. et al. Neuron 16, 1159–1167 (1996).

    Article  CAS  Google Scholar 

  13. Kohout, S.C. et al. Nat. Struct. Mol. Biol. 15, 106–108 (2008).

    Article  CAS  Google Scholar 

  14. McCobb, D.P. & Beam, K.G. Neuron 7, 119–127 (1991).

    Article  CAS  Google Scholar 

  15. Bean, B.P. Nat. Rev. Neurosci. 8, 451–465 (2007).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank M. Higa and staff at Sesoko Marine Station of Ryuku University for their support during our excursions for sample collection. This work was partly supported by grants from Japan Ministry of Education, Culture, Sports, Science and Technology Grant-in-Aid for Scientific Research on priority areas, the Human Frontier Science Program and Special Postdoctoral Researcher Program of RIKEN.

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Authors and Affiliations

  1. Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, 2-1 Hirosawa, Saitama, 351-0198, Japan

    Hidekazu Tsutsui, Satoshi Karasawa & Atsushi Miyawaki

  2. Amalgaam Co., Ltd. 2-9-3 Itabashi, Itabashi-ku, Tokyo, 173-0004, Japan

    Satoshi Karasawa

  3. Section of Developmental Neurophysiology, Okazaki Institute for Integrative Bioscience, Higashiyama 5-1, Myodaiji, Okazaki, Aichi, 444-8585, Japan

    Yasushi Okamura

Authors
  1. Hidekazu Tsutsui
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  2. Satoshi Karasawa
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  3. Yasushi Okamura
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  4. Atsushi Miyawaki
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Contributions

S.K. developed mKOκ. H.T. performed experiments to develop mUKG and Mermaid. H.T. and Y.O. designed experiments in regard to VSD. H.T. and A.M. designed experiments with regard to fluorescent proteins and FRET. H.T. and A.M. wrote the paper. A.M. supervised this project.

Corresponding author

Correspondence to Atsushi Miyawaki.

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Competing interests

S.K. is employed by Amalgaam. A.M. is a member of the scientific advisory board of Amalgaam. Amalgaam will sell mUKG and mKOκ

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6, Supplementary Methods, Supplementary Table 1 (PDF 868 kb)

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Tsutsui, H., Karasawa, S., Okamura, Y. et al. Improving membrane voltage measurements using FRET with new fluorescent proteins. Nat Methods 5, 683–685 (2008). https://doi.org/10.1038/nmeth.1235

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