Dual reporter approaches for identification of Cre efficacy and astrocyte heterogeneity

FASEB J. 2012 Nov;26(11):4576-83. doi: 10.1096/fj.12-207183. Epub 2012 Aug 2.

Abstract

Gene inactivation reporters are powerful tools to circumvent limitations of the widely used Cre/loxP system of conditional mutagenesis. With new conditional transgenic mouse lines expressing the enhanced cyan fluorescent protein (ECFP) instead of connexin43 (Cx43) after Cre-mediated recombination, we demonstrate dual reporter approaches to simultaneously examine astrocyte subpopulations expressing different connexins, identify compensatory up-regulation within gene families, and quantify Cre-mediated deletion at the allelic level. Analysis of a newly generated Cx43 knock-in ECFP mouse revealed an unexpected heterogeneity of Cx43-expressing astrocytes across brain areas.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Astrocytes / cytology
  • Astrocytes / metabolism*
  • Brain / metabolism
  • Connexin 30
  • Connexin 43 / genetics*
  • Connexin 43 / metabolism
  • Connexins / genetics*
  • Connexins / metabolism
  • Gene Deletion
  • Gene Expression Regulation / physiology*
  • Genes, Reporter*
  • Glial Fibrillary Acidic Protein
  • Green Fluorescent Proteins
  • Integrases / genetics
  • Integrases / metabolism*
  • Mice
  • Mice, Transgenic
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism

Substances

  • Connexin 30
  • Connexin 43
  • Connexins
  • Gjb6 protein, mouse
  • Glial Fibrillary Acidic Protein
  • Nerve Tissue Proteins
  • enhanced cyan fluorescent protein
  • glial fibrillary astrocytic protein, mouse
  • Green Fluorescent Proteins
  • Cre recombinase
  • Integrases