In vivo imaging and noninvasive ablation of pyramidal neurons in adult NEX-CreERT2 mice

Cereb Cortex. 2012 Jul;22(7):1473-86. doi: 10.1093/cercor/bhr214. Epub 2011 Aug 31.

Abstract

To study the function of individual neurons that are embedded in a complex neural network is difficult in mice. Conditional mutagenesis permits the spatiotemporal control of gene expression including the ablation of cells by toxins. To direct expression of a tamoxifen-inducible variant of Cre recombinase (CreERT2) selectively to cortical neurons, we replaced the coding region of the murine Nex1 gene by CreERT2 cDNA via homologous recombination in embryonic stem cells. When injected with tamoxifen, adult NEX-CreERT2 mice induced reporter gene expression exclusively in projection neurons of the neocortex and hippocampus. By titrating the tamoxifen dosage, we achieved recombination in single cells, which allowed multiphoton imaging of neocortical neurons in live mice. When hippocampal projection neurons were genetically ablated by induced expression of diphteria toxin, within 20 days the inflammatory response included the infiltration of CD3+ T cells. This marks a striking difference from similar studies, in which dying oligodendrocytes failed to recruit cells of the adaptive immune system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism*
  • Cell Survival / drug effects
  • Diphtheria Toxin / poisoning*
  • Genes, Reporter
  • Integrases / genetics
  • Integrases / metabolism*
  • Mice
  • Mice, Transgenic
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Poisons / pharmacology
  • Pyramidal Cells / cytology
  • Pyramidal Cells / drug effects
  • Pyramidal Cells / physiology*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*
  • Tamoxifen / pharmacology*

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • Diphtheria Toxin
  • Nerve Tissue Proteins
  • Neurod6 protein, mouse
  • Poisons
  • Recombinant Proteins
  • Tamoxifen
  • Cre recombinase
  • Integrases