B6.129P2(Cg)-Slc6a2^tm1.1Hhg/J

Stock number: 024166
Product name: NET-CI
Research areas: Cell Biology Research, Neurobiology Research

Description

NET-CI mice contain a triple mutation in the noradrenalin transporter gene rendering them insensitive to cocaine and antidepressants.

Detailed description

NET-CI mice contain the amino acid mutations F101C/A105G/N153T in exons 2 and 3 of the solute carrier family 6 (neurotransmitter transporter, noradrenalin), member 2 (Slc6a2) gene. Slc6a2 encodes the norepinephrine transporter (NET) which pumps the neurotransmitter norepinephrine out of the synapse back into cytosol. Norepinephrine release in the brain plays a major role in locomotor activity and reward behavior. Defects in norepinephrine transport reduces the rate of transport from the synapse, thus increasing the levels of norepinephrine in the brain. NET has been implicated in a number of norepinephrine-related disorders, including attention-deficit/hyperactivity disorder, depression and hypotension. Cocaine is able to block this transport, by binding directly to the NET, increasing locomotor-related norepinephrine activity. NET-CI mice contain a triple mutation in the transmembrane domain 2 (TM2) and TM3 of the norepinephrine transporter introducing a conformational change on the protein structure. These mice retain 75% functional NET activity and is rendered insensitive to cocaine. NET-CI NET is 37-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 2 of the solute carrier family 6 (neurotransmitter transporter, noradrenalin), member 2 (Slc6a2) gene. The double mutation, F101C/A105G, was introduced in exon 2 and a third mutation, N153T, was introduced into exon 3. This targeting construct was electroporated into 129P2/OlaHsd-derived E14TG2a.4 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 Sox2-cre expressing mice (see Stock No. 008454) to remove the floxed neo cassette. These NET-CI mice were bred to C57BL/6J mice for at least 5 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: Slc6a2^tm1.1Hhg
Name: targeted mutation 1.1, Haihua Gu
MGI accession ID: MGI:5547831
Generation method: Targeted
Synonyms: NET-CI
Marker symbol: Slc6a2
Marker name: solute carrier family 6 (neurotransmitter transporter, noradrenalin), member 2
Marker synonyms: NAT1; NE transporter; Net; NET1; norepinephrine transporter; SLC6A5
Chromosome: 8
Strain of origin: 129P2/OlaHsd
Molecular note: The targeting construct was designed to insert a loxP-flanked neomycin (neo) resistance cassette downstream of exon 2. The double mutation, F101C/A105G, was introduced in exon 2 and a third mutation, N153T, was introduced into exon 3. Cre-mediated recombination removed the floxed neo cassette.