B6.129X1(FVB)-Slc6a3^tm1Hhg/J

Stock number: 024165
Research areas: Cell Biology Research, Neurobiology Research

Description

DAT-CI mice contain a triple mutation in the dopamine transporter gene rendering them insensitive to cocaine.

Detailed description

DAT-CI mice contain the amino acid mutations L104V/F105C/A109V in exon 3 of the solute carrier family 6 (neurotransmitter transporter, dopamine), member 3 (Slc6a3) gene. Slc6a3 encodes the dopamine transporter (DAT) which pumps the neurotransmitter dopamine out of the synapse back into cytosol. Dopamine release in the ventral tegmental area (VTA) plays a major role in reward-motivated behavior. Defects in dopamine transport reduces the rate of transport from the synapse, thus increasing the levels of dopamine in the brain. DAT has been implicated in a number of dopamine-related disorders, including attention deficit hyperactivity disorder, bipolar disorder, clinical depression, and alcoholism. Cocaine is able to block this transport, by binding directly to the DAT, increasing reward-related dopamine activity. DAT-CI mice contain a triple mutation in the transmembrane domain 2 (TM2) of the dopamine transporter introducing a conformational change on the protein structure. These mice retain a functional DAT that is rendered insensitive to cocaine. DAT-CI DAT is 70-fold more insensitive to cocaine inhibition than WT mice.

Development

A targeting construct was designed to insert a loxP-flanked neomycin (neo) resistance cassette downstream of exon 3 of the solute carrier family 6 (neurotransmitter transporter, dopamine), member 3 (Slc6a3) gene. The triple mutation L104V/F105C/A109V was introduced in exon 3. This targeting construct was electroporated into 129X1/SvJ-derived embryonic stem (ES) cells and correctly targeted ES cells were injected into C57BL/6J blastocysts. The resulting chimeric males were bred to C57BL/6J females and offspring were subsequently bred to FVB/N-Tg(EIIa-cre)C5379Lmgd/J mice (Stock No. 003314) to remove the floxed neo cassette. These DAT-CI mice were bred to C57BL/6J mice for at least 20 generations. Upon arrival at The Jackson Laboratory, mutant mice were bred to C57BL/6J mice (Stock No. 000664) for at least one generation to establish the colony.

Allele

Symbol: Slc6a3^tm1Hhg
Name: targeted mutation 1, Howard H Gu
MGI accession ID: MGI:3653596
Generation method: Targeted
Attributes: Hypomorph; Inserted expressed sequence
Marker symbol: Slc6a3
Marker name: solute carrier family 6 (neurotransmitter transporter, dopamine), member 3
Chromosome: 13
Strain of origin: 129X1/SvJ
Molecular note: A loxP-flanked neo was inserted into intron 3 with the L104V, F105C, and A109V mutations. The neo was subsequently removed from the locus.