STOCK Npy2r^{tm1.1(cre)Lbrl}/RcngJ

Stock number: 029285
Product name: Npy2r-ires-Cre
Research areas: Internal/Organ Research, Neurobiology Research, Research Tools

Description

Npy2r-ires-Cre knock-in mice express Cre recombinase under the direction of the Npy2r promoter in vagal sensory neurons in the lung and brain stem that control respiration.

Detailed description

Npy2r-ires-Cre mice have an IRES-Cre fusion protein inserted 3 base pairs downstream of the stop codon of the neuropeptide Y receptor Y2 (Npy2r) gene. The presence of IRES-Cre cassette downstream of the stop codon allows for expression of NPY2R and cre. NPY2R encodes a receptor present on many cell types, including a subset of slow-conducting unmyelinated A fiber vagal sensory neurons in the lung and brain stem. The Npy2r-ires-Cre reporter is expressed in 94% of Npy2r neurons when bred to a cre-dependent EYFP reporter. Homozygous mice are viable and fertile. When these mice are bred with mice containing loxP-flanked sequence, Cre-mediated recombination will result in deletion of the floxed sequences in the Cre-expressing cells of the offspring.

When Npy2r-ires-Cre mice were bred to B6;129S-Gt(ROSA)26Sor^{tm32(CAG-COP4*H134R/EYFP)Hze}/J (Stock No. 012569) mice, which express a cre-inducible improved channelrhodopsin-2/EYFP fusion protein, optogenetic activation of NPY2R neurons results in rapid and shallow breathing with decreases in heart rate and gastric pressure, which are also under vagal control. Activation causes a 68% increase in respiration rate, a 44% decrease in tidal volume, and a 3% decrease in minute volume.

 

Development

A targeting construct was designed by Stephen Liberles (Harvard Medical School) to insert an internal ribosomal entry site (IRES) fused to Cre Recombinase with a polyadenylation signal, followed by a frt-flanked neomycin resistance (neo) cassette, 3 base pairs downstream of the stop codon of the neuropeptide Y receptor Y2 (Npy2r) gene. This construct was electroporated into 129S6/SvEvTac-derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to B6;SJL-Tg(ACTFLPe)9205Dym/J mice (Stock No. 003800) to remove the neo cassette. Offspring were crossed to remove the flpe-expressing transgene, and resulting Npy2r-ires-Cre mice were bred to C57BL/6J mice for at least 1 generation. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Npy2r^{tm1.1(cre)Lbrl}
Name: targeted mutation 1.1, Stephen Liberles
MGI accession ID: MGI:5776535
Generation method: Targeted
Attributes: Recombinase-expressing
Synonyms: Npy2r-ires-Cre
Marker symbol: Npy2r
Marker name: neuropeptide Y receptor Y2
Marker synonyms: NPY2-R; NPY-Y2 receptor
Chromosome: 3
Strain of origin: 129S6/SvEvTac
Expressed genes: cre,cre recombinase,bacteriophage P1
Site of expression: Cre recombinase is expressed in vagal sensory neurons in the lung and brain stem.
Molecular note: A cassette encoding IRES, Cre recombinase, and an FRT-flanked neomycin selection cassette is inserted into the mouse Npy2r locus 3 base pairs downstream of the coding sequence stop codon.