title: Imaging activity in neurons and glia with a Polr2a-based and cre-dependent GCaMP5G-IRES-tdTomato reporter mouse.
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.
authors.affiliation: 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.
authors.forename: J Michael
authors.initials: JM
authors.lastname: Gee
authors.affiliation: 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.
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Smith
authors.affiliation: Department of Bioengineering, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
authors.forename: Fernando R
authors.initials: FR
authors.lastname: Fernandez
authors.affiliation: Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
authors.forename: Michael N
authors.initials: MN
authors.lastname: Economo
authors.affiliation: Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
authors.forename: Daniela
authors.initials: D
authors.lastname: Brunert
authors.affiliation: Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
authors.forename: Markus
authors.initials: M
authors.lastname: Rothermel
authors.affiliation: Department of Bioengineering, University of Utah, Salt Lake City, UT 84112.
authors.forename: S Craig
authors.initials: SC
authors.lastname: Morris
authors.affiliation: Department of Human Genetics, University of Utah, Salt Lake City, UT 84112.
authors.forename: Amy
authors.initials: A
authors.lastname: Talbot
authors.affiliation: Department of Human Genetics, University of Utah, Salt Lake City, UT 84112.
authors.forename: Sierra
authors.initials: S
authors.lastname: Palumbos
authors.affiliation: Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Ichida
authors.affiliation: Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112.
authors.forename: Jason D
authors.initials: JD
authors.lastname: Shepherd
authors.affiliation: Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, UT 84112.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: West
authors.affiliation: Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
authors.forename: Matt
authors.initials: M
authors.lastname: Wachowiak
authors.affiliation: Department of Human Genetics, University of Utah, Salt Lake City, UT 84112; Howard Hughes Medical Institute, University of Utah, Salt Lake City, UT 84112.
authors.forename: Mario R
authors.initials: MR
authors.lastname: Capecchi
authors.affiliation: Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, UT 84112.
authors.forename: Karen S
authors.initials: KS
authors.lastname: Wilcox
authors.affiliation: 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.
authors.forename: John A
authors.initials: JA
authors.lastname: White
authors.affiliation: Department of Human Genetics, University of Utah, Salt Lake City, UT 84112. Electronic address: petr.tvrdik@genetics.utah.edu.
authors.forename: Petr
authors.initials: P
authors.lastname: Tvrdik
mesh_terms: D000344:Afferent Pathways / Q000502:physiology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002540:Cerebral Cortex / Q000502:physiology; D005260:Female; D017930:Genes, Reporter / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D066298:In Vitro Techniques; D019426:Integrases; D008297:Male; D008564:Membrane Potentials / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009457:Neuroglia / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D018408:Patch-Clamp Techniques; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D022661:RNA, Untranslated / Q000235:genetics / Q000378:metabolism; D014738:Vibrissae / Q000294:innervation
chemical_list: C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D022661:RNA, Untranslated; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases; C500816:POLR2B protein, human; D012319:RNA Polymerase II; D002118:Calcium