Imaging Activity in Neurons and Glia with a Polr2a-Based and Cre-Dependent GCaMP5G-IRES-tdTomato Reporter Mouse

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 locu...

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Bibliographic Details
Published in:	Neuron (Cambridge, Mass.) Mass.), 2014-09, Vol.83 (5), p.1058-1072
Main Authors:	Gee, J. Michael, Smith, Nathan A., Fernandez, Fernando R., Economo, Michael N., Brunert, Daniela, Rothermel, Markus, Morris, S. Craig, Talbot, Amy, Palumbos, Sierra, Ichida, Jennifer M., Shepherd, Jason D., West, Peter J., Wachowiak, Matt, Capecchi, Mario R., Wilcox, Karen S., White, John A., Tvrdik, Petr
Format:	Article
Language:	eng
Subjects:	Afferent Pathways - physiology Animals Calcium - metabolism Cerebral Cortex - physiology Female Genes, Reporter - physiology Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism In Vitro Techniques Integrases Male Membrane Potentials - genetics Mice Mice, Inbred C57BL Mice, Transgenic Neuroglia - physiology Neurons - physiology Patch-Clamp Techniques RNA Polymerase II - genetics RNA Polymerase II - metabolism RNA, Untranslated - genetics RNA, Untranslated - metabolism Vibrissae - innervation
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Summary:	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. [Display omitted] •PC::G5-tdT is a versatile Cre-dependent mouse reporter of intracellular calcium•The reporter is based on the GCaMP5G indicator targeted to the Polr2a locus•PC::G5-tdT reveals calcium activity in neuronal as well as glial somas and processes•Inclusion of a tdTomato tag facilitates monitoring of expression patterns Gee et al. developed a calcium reporter mouse for imaging neural activity in vivo. The reporter detects activity in somas and processes of neurons, astrocytes, and microglia. It is a promising tool for monitoring neuronal-glial interactions in neurological models.
ISSN:	0896-6273 1097-4199