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Imaging activity in neurons and glia with a Polr2a-based and cre-dependent GCaMP5G-IRES-tdTomato reporter mouse.
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.
Afiliação
  • Gee JM; 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.
  • Smith NA; 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.
  • Fernandez FR; Department of Bioengineering, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
  • Economo MN; Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
  • Brunert D; Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
  • Rothermel M; Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
  • Morris SC; Department of Bioengineering, University of Utah, Salt Lake City, UT 84112.
  • Talbot A; Department of Human Genetics, University of Utah, Salt Lake City, UT 84112.
  • Palumbos S; Department of Human Genetics, University of Utah, Salt Lake City, UT 84112.
  • Ichida JM; Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112.
  • Shepherd JD; Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112.
  • West PJ; Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, UT 84112.
  • Wachowiak M; Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, UT 84112; Brain Institute, University of Utah, Salt Lake City, UT 84112.
  • Capecchi MR; Department of Human Genetics, University of Utah, Salt Lake City, UT 84112; Howard Hughes Medical Institute, University of Utah, Salt Lake City, UT 84112.
  • Wilcox KS; Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, UT 84112.
  • White JA; 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.
  • Tvrdik P; Department of Human Genetics, University of Utah, Salt Lake City, UT 84112. Electronic address: petr.tvrdik@genetics.utah.edu.
Neuron ; 83(5): 1058-72, 2014 Sep 03.
Article em En | MEDLINE | ID: mdl-25155958
ABSTRACT
New strategies for introducing genetically encoded activity indicators into animal models facilitate the investigation of nervous system function. We have developed the PCG5-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, PCG5-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. PCG5-tdT will be instrumental for a variety of brain mapping experiments.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: RNA Polimerase II / Neuroglia / Cálcio / Genes Reporter / Neurônios Tipo de estudo: Prognostic_studies Limite: Animals Idioma: En Ano de publicação: 2014 Tipo de documento: Article
Texto completo
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: RNA Polimerase II / Neuroglia / Cálcio / Genes Reporter / Neurônios Tipo de estudo: Prognostic_studies Limite: Animals Idioma: En Ano de publicação: 2014 Tipo de documento: Article