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

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.

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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).

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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).

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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.

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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.

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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

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