A noninvasive genetic/pharmacologic strategy for visualizing cell morphology and clonal relationships in the mouse

J Neurosci. 2003 Mar 15;23(6):2314-22. doi: 10.1523/JNEUROSCI.23-06-02314.2003.

Abstract

Analysis of cellular morphology is the most general approach to neuronal classification. With the increased use of genetically engineered mice, there is a growing need for methods that can selectively visualize the morphologies of specified subsets of neurons. This capability is needed both to define cell morphologic phenotypes and to mark cells in a noninvasive manner for lineage studies. To this end, we describe a bipartite genetic system based on a Cre-estrogen receptor (ER) fusion protein that irreversibly activates a plasma membrane-bound alkaline phosphatase reporter gene by site-specific recombination. Because the efficiency and timing of gene rearrangement is controlled pharmacologically, a sparse subset of labeled cells can be generated from the set of CreER-expressing cells at any time during development. Histochemical visualization of alkaline phosphatase activity reveals neuronal morphology with strong and uniform labeling of all processes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / analysis
  • Alkaline Phosphatase / metabolism
  • Animals
  • Brain / cytology
  • Brain / growth & development
  • Cell Lineage / physiology
  • Clone Cells / classification*
  • Clone Cells / cytology*
  • Clone Cells / metabolism
  • Gene Targeting
  • Genes, Reporter
  • Histocytochemistry / methods*
  • Integrases / genetics
  • Mice
  • Neurons / classification*
  • Neurons / cytology*
  • Neurons / metabolism
  • Phenotype
  • Receptors, Estrogen / genetics
  • Recombinant Fusion Proteins / drug effects
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombination, Genetic
  • Reproducibility of Results
  • Selective Estrogen Receptor Modulators / pharmacology
  • Tamoxifen / pharmacology
  • Transgenes
  • Viral Proteins / genetics

Substances

  • Receptors, Estrogen
  • Recombinant Fusion Proteins
  • Selective Estrogen Receptor Modulators
  • Viral Proteins
  • Tamoxifen
  • Cre recombinase
  • Integrases
  • Alkaline Phosphatase