Analysis of dopamine transporter gene expression pattern -- generation of DAT-iCre transgenic mice.
The dopamine transporter is an essential component of the dopaminergic synapse. It is located in the presynaptic neurons and regulates extracellular dopamine levels. We generated a transgenic mouse line expressing the Cre recombinase under the control of the regulatory elements of the dopamine transporter gene, for investigations of gene function in dopaminergic neurons. The codon-improved Cre recombinase (iCre) gene was inserted into the dopamine transporter gene on a bacterial artificial chromosome. The pattern of expression of the bacterial artificial chromosome-dopamine transporter-iCre transgene was similar to that of the endogenous dopamine transporter gene, as shown by immunohistochemistry. Recombinase activity was further studied in mice carrying both the bacterial artificial chromosome-dopamine transporter-iCre transgene and a construct expressing the beta-galactosidase gene after Cre-mediated recombination. In situ studies showed that beta-galactosidase (5-bromo-4-chloroindol-3-yl beta-D-galactoside staining) and the dopamine transporter (immunofluorescence) had identical distributions in the ventral midbrain. We used this animal model to study the distribution of dopamine transporter gene expression in hypothalamic nuclei in detail. The expression profile of tyrosine hydroxylase (an enzyme required for dopamine synthesis) was broader than that of beta-galactosidase in A12 to A15. Thus, only a fraction of neurons synthesizing dopamine expressed the dopamine transporter gene. The bacterial artificial chromosome-dopamine transporter-iCre transgenic line is a unique tool for targeting Cre/loxP-mediated DNA recombination to dopamine neurons for studies of gene function or for labeling living cells, following the crossing of these mice with transgenic Cre reporter lines producing fluorescent proteins.
Turiault
Marc
M
 CNRS UMR7148, Molecular Genetics, Neurophysiology and Behavior, Collège de France, Institut de Biologie, Paris, France Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany.
Parnaudeau
Sébastien
S
 CNRS UMR7148, Molecular Genetics, Neurophysiology and Behavior, Collège de France, Institut de Biologie, Paris, France Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany.
Milet
Aude
A
 CNRS UMR7148, Molecular Genetics, Neurophysiology and Behavior, Collège de France, Institut de Biologie, Paris, France Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany.
Parlato
Rosanna
R
 CNRS UMR7148, Molecular Genetics, Neurophysiology and Behavior, Collège de France, Institut de Biologie, Paris, France Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany.
Rouzeau
Jean-Denis
JD
 CNRS UMR7148, Molecular Genetics, Neurophysiology and Behavior, Collège de France, Institut de Biologie, Paris, France Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany.
Lazar
Monique
M
 CNRS UMR7148, Molecular Genetics, Neurophysiology and Behavior, Collège de France, Institut de Biologie, Paris, France Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany.
Tronche
François
F
 CNRS UMR7148, Molecular Genetics, Neurophysiology and Behavior, Collège de France, Institut de Biologie, Paris, France Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany.
D000818:Animals; D001522:Behavior, Animal; D050483:Dopamine Plasma Membrane Transport Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000235:genetics*; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009043:Motor Activity; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D050483:Dopamine Plasma Membrane Transport Proteins; D011994:Recombinant Proteins; C045073:Cre recombinase; D019426:Integrases; D001616:beta-Galactosidase