Format

Send to

Choose Destination
See comment in PubMed Commons below
Neuron. 2014 Sep 3;83(5):1058-72. doi: 10.1016/j.neuron.2014.07.024. Epub 2014 Aug 21.

Imaging activity in neurons and glia with a Polr2a-based and cre-dependent GCaMP5G-IRES-tdTomato reporter mouse.

Author information

1
Department of Bioengineering, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112; MD-PhD Program, University of Utah, Salt Lake City, UT 84112.
2
Department of Bioengineering, University of Utah, Salt Lake City, UT 84112; Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
3
Department of Bioengineering, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
4
Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
5
Department of Bioengineering, University of Utah, Salt Lake City, UT 84112.
6
Department of Human Genetics, University of Utah, Salt Lake City, UT 84112.
7
Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112.
8
Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, UT 84112.
9
Department of Human Genetics, University of Utah, Salt Lake City, UT 84112; Howard Hughes Medical Institute, University of Utah, Salt Lake City, UT 84112.
10
Department of Bioengineering, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112. Electronic address: john.white@utah.edu.
11
Department of Human Genetics, University of Utah, Salt Lake City, UT 84112. Electronic address: petr.tvrdik@genetics.utah.edu.

Abstract

New strategies for introducing genetically encoded activity indicators into animal models facilitate the investigation of nervous system function. We have developed the PC::G5-tdT mouse line that expresses the GCaMP5G calcium indicator in a Cre-dependent fashion. Instead of targeting the ROSA26 locus, we inserted the reporter cassette nearby the ubiquitously expressed Polr2a gene without disrupting locus integrity. The indicator was tagged with IRES-tdTomato to aid detection of positive cells. This reporter system is effective in a wide range of developmental and cellular contexts. We recorded spontaneous cortical calcium waves in intact awake newborns and evaluated concentration-dependent responses to odorants in the adult olfactory bulb. Moreover, PC::G5-tdT effectively reports intracellular calcium dynamics in somas and fine processes of astrocytes and microglial cells. Through electrophysiological and behavioral analyses, we determined that GCaMP5G expression had no major impact on nervous system performance. PC::G5-tdT will be instrumental for a variety of brain mapping experiments.

VIDEO ABSTRACT:

PMID:
25155958
PMCID:
PMC4156920
DOI:
10.1016/j.neuron.2014.07.024
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science Icon for PubMed Central
    Loading ...
    Support Center