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

Neuron. 2014 Sep 3;83(5):1058-72. doi: 10.1016/j.neuron.2014.07.024. Epub 2014 Aug 21.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • 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

Substances

  • Gt(ROSA)26Sor non-coding RNA, mouse
  • RNA, Untranslated
  • Green Fluorescent Proteins
  • Cre recombinase
  • Integrases
  • POLR2B protein, human
  • RNA Polymerase II
  • Calcium