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PLoS One. 2017 Jun 22;12(6):e0179460. doi: 10.1371/journal.pone.0179460. eCollection 2017.

An R-CaMP1.07 reporter mouse for cell-type-specific expression of a sensitive red fluorescent calcium indicator.

Author information

1
Brain Research Institute, University of Zurich, Zurich, Switzerland.
2
Neuroscience Center Zurich (ZNZ), University of Zurich and ETH Zurich, Zurich, Switzerland.
3
Institute of Neuropathology, University Hospital of Zurich, Zurich, Switzerland.
4
Allen Institute for Brain Science, Seattle, Washington, United States of America.
5
Brain Mind Institute, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
6
Saitama University, Brain Science Institute, Saitama, Japan.

Abstract

Genetically encoded calcium indicators (GECIs) enable imaging of in vivo brain cell activity with high sensitivity and specificity. In contrast to viral infection or in utero electroporation, indicator expression in transgenic reporter lines is induced noninvasively, reliably, and homogenously. Recently, Cre/tTA-dependent reporter mice were introduced, which provide high-level expression of green fluorescent GECIs in a cell-type-specific and inducible manner when crossed with Cre and tTA driver mice. Here, we generated and characterized the first red-shifted GECI reporter line of this type using R-CaMP1.07, a red fluorescent indicator that is efficiently two-photon excited above 1000 nm. By crossing the new R-CaMP1.07 reporter line to Cre lines driving layer-specific expression in neocortex we demonstrate its high fidelity for reporting action potential firing in vivo, long-term stability over months, and versatile use for functional imaging of excitatory neurons across all cortical layers, especially in the previously difficult to access layers 4 and 6.

PMID:
28640817
PMCID:
PMC5480891
DOI:
10.1371/journal.pone.0179460
[Indexed for MEDLINE]
Free PMC Article

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