Spontaneous network activity visualized by ultrasensitive Ca2+ indicators, yellow Cameleon-Nano (original) (raw)

Nature Methods volume 7, pages 729–732 (2010)Cite this article

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Abstract

We report ultrasensitive Ca2+ indicators, yellow cameleon-Nano (YC-Nano), developed by engineering the Ca2+-sensing domain of a genetically encoded Ca2+ indicator, YC2.60 or YC3.60. Their high Ca2+ affinities (_K_d = 15–140 nM) and large signal change (1,450%) enabled detection of subtle Ca2+ transients associated with intercellular signaling dynamics and neuronal activity, even in 100,000-cell networks. These indicators will be useful for studying information processing in living multicellular networks.

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References

  1. Paredes, R.M., Etzler, J.C., Watts, L.T., Zheng, W. & Lechleiter, J.D. Methods 46, 143–151 (2008).
    Article CAS Google Scholar
  2. Kotlikoff, M.I. J. Physiol. (Lond.) 578, 55–67 (2007).
    Article CAS Google Scholar
  3. Stosiek, C., Garaschuk, O., Holthoff, K. & Konnerth, A. Proc. Natl. Acad. Sci. USA 100, 7319–7324 (2003).
    Article CAS Google Scholar
  4. Wallace, D.J. et al. Nat. Methods 5, 797–804 (2008).
    Article CAS Google Scholar
  5. Mank, M. et al. Nat. Methods 5, 805–811 (2008).
    Article CAS Google Scholar
  6. Tian, L. et al. Nat. Methods 6, 875–881 (2009).
    Article CAS Google Scholar
  7. Abe, T., Maeda, Y. & Iijima, T. Differentiation 39, 90–96 (1988).
    Article CAS Google Scholar
  8. Zhang, W.H., Rengel, Z. & Kuo, J. Plant J. 15, 147–151 (1998).
    Article Google Scholar
  9. Hendel, T. et al. J. Neurosci. 28, 7399–7411 (2008).
    Article CAS Google Scholar
  10. Yasuda, R. et al. Sci. STKE 2004, pl5 (2004).
    Google Scholar
  11. Palmer, A.E. & Tsien, R.Y. Nat. Protoc. 1, 1057–1065 (2006).
    Article CAS Google Scholar
  12. Nagai, T., Yamada, S., Tominaga, T., Ichikawa, M. & Miyawaki, A. Proc. Natl. Acad. Sci. USA 101, 10554–10559 (2004).
    Article CAS Google Scholar
  13. Miyawaki, A. et al. Nature 388, 882–887 (1997).
    Article CAS Google Scholar
  14. Martin, S.R. et al. Biochemistry 35, 3508–3517 (1996).
    Article CAS Google Scholar
  15. Porumb, T., Yau, P., Harvey, T.S. & Ikura, M. Protein Eng. 7, 109–115 (1994).
    Article CAS Google Scholar
  16. Tsien, R.Y. & Pozzan, T. Methods Enzymol. 172, 230–262 (1989).
    Article CAS Google Scholar
  17. Bers, D.M. Am. J. Physiol. 242, C404–C408 (1982).
    Article CAS Google Scholar
  18. Matsu-ura, T. et al. J. Cell Biol. 173, 755–765 (2006).
    Article CAS Google Scholar
  19. Nagasaki, A., de Hostos, E.L. & Uyeda, T.Q.P. J. Cell Sci. 115, 2241–2251 (2001).
    Google Scholar
  20. Mizuguchi, H. & Kay, M.A. Hum. Gene Ther. 9, 2577–2583 (1998).
    Article CAS Google Scholar
  21. Fukuda, H., Terashima, M., Koshikawa, M., Kanegae, Y. & Saito, I. Microbiol. Immunol. 50, 643–654 (2006).
    Article CAS Google Scholar
  22. Davie, J.T. et al. Nat. Protoc. 1, 1235–1247 (2006).
    Article CAS Google Scholar
  23. Westerfield, M. The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio). 4th edn. (Univ. of Oregon Press, Eugene, 2000).

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Acknowledgements

We thank A. Nagasaki and Y. Kuramoto for instruction and assistance with the experiment using Dictyostelium cell, T. Kotani and S. Higashijima for assistance and instruction on fish embryo imaging, and T. Shimogori for instruction on in utero injection. This work was partly supported by a Grant-in-Aid for Young Scientists (A) of the Japan Society for the Promotion of Science and Scientific Research on Advanced Medical Technology of the Ministry of Labor, Health and Welfare of Japan to T.N. and a grant from Precursory Research for Embryonic Science and Technology of the Japan Science and Technology Agency to T.N. and K.H.

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Author notes

  1. Kazuki Horikawa and Yoshiyuki Yamada: These authors contributed equally to this work.

Authors and Affiliations

  1. Research Institute for Electronic Science, Hokkaido University, Sapporo, Hokkaido, Japan
    Kazuki Horikawa, Tomoki Matsuda, Kentarou Kobayashi & Takeharu Nagai
  2. Precursory Research for Embryonic Science and Technology, Japan Science and Technology Agency, Tokyo, Japan
    Kazuki Horikawa & Takeharu Nagai
  3. Japan Science and Technology Agency, International Cooperative Research Project and Solution-Oriented Research for Science and Technology, Calcium Oscillation Project, Saitama, Japan
    Yoshiyuki Yamada, Takayuki Michikawa & Katsuhiko Mikoshiba
  4. Laboratory for Developmental Neurobiology, RIKEN Brain Science Institute, Saitama, Japan
    Yoshiyuki Yamada, Toru Matsu-ura, Takayuki Michikawa & Katsuhiko Mikoshiba
  5. Hashimoto Research Unit, RIKEN Brain Science Institute, Saitama, Japan
    Mitsuhiro Hashimoto
  6. Laboratory for Cell Function Dynamics, RIKEN Brain Science Institute, Saitama, Japan
    Atsushi Miyawaki

Authors

  1. Kazuki Horikawa
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  2. Yoshiyuki Yamada
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  3. Tomoki Matsuda
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  4. Kentarou Kobayashi
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  5. Mitsuhiro Hashimoto
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  6. Toru Matsu-ura
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  7. Atsushi Miyawaki
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  8. Takayuki Michikawa
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  9. Katsuhiko Mikoshiba
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  10. Takeharu Nagai
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Contributions

K.H. and T.N. invented YC-Nano variants. Y.Y., M.H., A.M., T. Michikawa and K.M. established the method of the expression of Ca2+ indicators in neurons using adenoviral vectors; K.H. and T. Matsuda performed experiments other than electrophysiology and Ca2+ imaging in brain slices. K.H. and T. Matsu-ura performed stopped-flow spectrometry. Y.Y. performed electrophysiology and Ca2+ imaging in brain slices; K.H., K.K., Y.Y., T. Michikawa and T.N. analyzed data. K.H., Y.Y., T. Michikawa and T.N. wrote the manuscript. T.N. supervised the study.

Corresponding author

Correspondence toTakeharu Nagai.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7, Supplementary Tables 1–2 and Supplementary Notes 1–5 (PDF 1898 kb)

Supplementary Video 1

Comparative visualization of Ca2+ dynamics in early aggregating stage of Dictyostelium cells visualized by YC-Nano15 and YC2.60. (MOV 908 kb)

Supplementary Video 2

Ca2+ dynamics in aggregating Dictyostelium cells visualized by YC-Nano15. (MOV 1606 kb)

Supplementary Video 3

Ca2+ dynamics in aggregating Dictyostelium cells visualized by YC2.60. (MOV 224 kb)

Supplementary Video 4

Twitching behavior of zebrafish embryo. (MOV 196 kb)

Supplementary Video 5

Spontaneous motor activities in living zebrafish embryos visualized by YC-Nano50. (MOV 2568 kb)

Supplementary Video 6

Spontaneous motor activities in living zebrafish embryos visualized by YC3.60. (MOV 1520 kb)

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Horikawa, K., Yamada, Y., Matsuda, T. et al. Spontaneous network activity visualized by ultrasensitive Ca2+ indicators, yellow Cameleon-Nano.Nat Methods 7, 729–732 (2010). https://doi.org/10.1038/nmeth.1488

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