Targeted gene expression in dopamine and serotonin neurons of the mouse brain.
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.
Zhuang
Xiaoxi
X
Department of Neurobiology, Pharmacology and Physiology, The University of Chicago, 924 East 57th Street, Knapp Center, R214, Chicago, IL 60637, USA. xzhuang@bsd.uchicago.edu
Masson
Justine
J
Gingrich
Jay A
JA
Rayport
Stephen
S
Hen
René
R
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002678:Chimera; D004298:Dopamine / Q000096:biosynthesis*; D050483:Dopamine Plasma Membrane Transport Proteins; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology; D018390:Gene Targeting / Q000379:methods; D017930:Genes, Reporter / Q000235:genetics; D019426:Integrases / Q000235:genetics; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics*; D026901:Membrane Transport Proteins / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008967:Molecular Biology / Q000379:methods; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009448:Neurochemistry / Q000379:methods; D009474:Neurons / Q000737:chemistry / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012701:Serotonin / Q000096:biosynthesis*; D050486:Serotonin Plasma Membrane Transport Proteins; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D050483:Dopamine Plasma Membrane Transport Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; D050486:Serotonin Plasma Membrane Transport Proteins; C493215:Slc6a3 protein, mouse; C493231:Slc6a4 protein, mouse; D012701:Serotonin; D014446:Tyrosine 3-Monooxygenase; C045073:Cre recombinase; D019426:Integrases; D001616:beta-Galactosidase; D004298:Dopamine