Skip to main content
Log in

Improved calcium imaging in transgenic mice expressing a troponin C–based biosensor

  • Brief Communication
  • Published:

From Nature Methods

View current issue Submit your manuscript

Abstract

Fluorescent Ca2+ indicator proteins (FCIPs) are attractive tools for studying Ca2+ dynamics in live cells. Here we describe transgenic mouse lines expressing a troponin C (TnC)-based biosensor. The biosensor is widely expressed in neurons and has improved Ca2+ sensitivity both in vitro and in vivo. This allows FCIP-based two-photon Ca2+ imaging of distinct neurons and their dendrites in vivo, and opens a new avenue for structure-function analysis of intact neuronal circuits.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Figure 1: Characterization of Thy1-CerTN-L15 transgenic mice.
Figure 2: Ca2+ sensitivity of CerTN-L15.
Figure 3: CerTN-L15–based Ca2+ measurements in vivo. (a) Micrograph of a spiny dendrite located 30 μm below the cortical surface (top).

Similar content being viewed by others

References

  1. Zhang, J., Campbell, R.E., Ting, A.Y. & Tsien, R.Y. Nat. Rev. Mol. Cell Biol. 3, 906–918 (2002).

    CAS  PubMed  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  3. Nagai, T., Yamada, S., Tominaga, T., Ichikawa, M. & Miyawaki, A. Proc. Natl. Acad. Sci. USA 101, 10554–10559 (2004).

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Hasan, M.T. et al. PLoS Biol. 2, 763–775 (2004).

    CAS  Google Scholar 

  5. Diez-Garcia, J. et al. Eur. J. Neurosci. 22, 627–635 (2005).

    PubMed  Google Scholar 

  6. Heim, N. & Griesbeck, O. J. Biol. Chem. 279, 14280–14286 (2004).

    CAS  PubMed  Google Scholar 

  7. Rizzo, M.A., Springer, G.H., Granada, B. & Piston, D.W. Nat. Biotechnol. 22, 445–449 (2004).

    CAS  PubMed  Google Scholar 

  8. Griesbeck, O., Baird, G.S., Campbell, R.E., Zacharias, D.A. & Tsien, R.Y. J. Biol. Chem. 276, 29188–29194 (2001).

    CAS  PubMed  Google Scholar 

  9. Caroni, P. J. Neurosci. Methods 71, 3–9 (1997).

    CAS  PubMed  Google Scholar 

  10. Jurado, L.A., Chockalingam, P.S. & Jarrett, H.W. Physiol. Rev. 79, 661–682 (1999).

    CAS  PubMed  Google Scholar 

  11. Benaim, G. & Villalobo, A. Eur. J. Biochem. 269, 3619–3631 (2002).

    CAS  PubMed  Google Scholar 

  12. Garaschuk, O., Milos, R.I. & Konnerth, A. Nat. Protocols 1, 380–386 (2006).

    CAS  PubMed  Google Scholar 

  13. Stosiek, C., Garaschuk, O., Holthoff, K. & Konnerth, A. Proc. Natl. Acad. Sci. USA 100, 7319–7324 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank M. Bösl for doing pronucleus injections, and A. Schulze-Schenke and S. Direnberger for assistance. Supported by grants of the Deutsche Forschungsgemeinschaft (SFB 391 and SFB 596) and the Bundesministerium für Bildung und Forschung (NGFN-2).

Author information

Authors and Affiliations

Authors

Contributions

N.H., M.W.F., M.M. and O.G. generated transgenic mice and performed biochemical and spectroscopic characterization; O.Ga., R.I.M., Y.K. and A.K. performed electrophysiology and two-photon imaging.

Corresponding authors

Correspondence to Arthur Konnerth or Oliver Griesbeck.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

In vitro characterization of CerTN-L15.

Supplementary Fig. 2

Expression pattern of CerTN-L15 in the brain of a Thy 1-CerTN-L15 transgenic mouse.

Supplementary Fig. 3

In vivo neuronal and astrocytic morphology in the cortex of a Thy 1-CerTN-L15 transgenic mouse.

Supplementary Table 1

Sensor expression patterns in three Thy 1-CerTN-L15 mouse lines with low, medium and high expression.

Supplementary Methods

Rights and permissions

Reprints and permissions

About this article

Cite this article

Heim, N., Garaschuk, O., Friedrich, M. et al. Improved calcium imaging in transgenic mice expressing a troponin C–based biosensor. Nat Methods 4, 127–129 (2007). https://doi.org/10.1038/nmeth1009

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth1009

  • Springer Nature America, Inc.

This article is cited by

Navigation