Targeted gene expression in dopamine and serotonin neurons of the mouse brain

J Neurosci Methods. 2005 Apr 15;143(1):27-32. doi: 10.1016/j.jneumeth.2004.09.020. Epub 2004 Nov 24.

Abstract

We used a knock-in strategy to generate two lines of mice expressing Cre recombinase under the transcriptional control of the dopamine transporter promoter (DAT-cre mice) or the serotonin transporter promoter (SERT-cre mice). In DAT-cre mice, immunocytochemical staining of adult brains for the dopamine-synthetic enzyme tyrosine hydroxylase and for Cre recombinase revealed that virtually all dopaminergic neurons in the ventral midbrain expressed Cre. Crossing DAT-cre mice with ROSA26-stop-lacZ or ROSA26-stop-YFP reporter mice revealed a near perfect correlation between staining for tyrosine hydroxylase and beta-galactosidase or YFP. YFP-labeled fluorescent dopaminergic neurons could be readily identified in live slices. Crossing SERT-cre mice with the ROSA26-stop-lacZ or ROSA26-stop-YFP reporter mice similarly revealed a near perfect correlation between staining for serotonin-synthetic enzyme tryptophan hydroxylase and beta-galactosidase or YFP. Additional Cre expression in the thalamus and cortex was observed, reflecting the known pattern of transient SERT expression during early postnatal development. These findings suggest a general strategy of using neurotransmitter transporter promoters to drive selective Cre expression and thus control mutations in specific neurotransmitter systems. Crossed with fluorescent-gene reporters, this strategy tags neurons by neurotransmitter status, providing new tools for electrophysiology and imaging.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Brain / metabolism*
  • Chimera
  • Dopamine / biosynthesis*
  • Dopamine Plasma Membrane Transport Proteins
  • Female
  • Gene Expression Regulation / physiology
  • Gene Targeting / methods
  • Genes, Reporter / genetics
  • Integrases / genetics
  • Male
  • Membrane Glycoproteins / genetics*
  • Membrane Transport Proteins / genetics*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Molecular Biology / methods
  • Nerve Tissue Proteins / genetics*
  • Neurochemistry / methods
  • Neurons / chemistry
  • Neurons / metabolism*
  • Promoter Regions, Genetic / genetics
  • Recombinant Fusion Proteins / genetics
  • Serotonin / biosynthesis*
  • Serotonin Plasma Membrane Transport Proteins
  • Tyrosine 3-Monooxygenase / genetics
  • beta-Galactosidase / genetics

Substances

  • Dopamine Plasma Membrane Transport Proteins
  • Membrane Glycoproteins
  • Membrane Transport Proteins
  • Nerve Tissue Proteins
  • Recombinant Fusion Proteins
  • Serotonin Plasma Membrane Transport Proteins
  • Slc6a3 protein, mouse
  • Slc6a4 protein, mouse
  • Serotonin
  • Tyrosine 3-Monooxygenase
  • Cre recombinase
  • Integrases
  • beta-Galactosidase
  • Dopamine