B6.129S4-Agrp^{tm2(HBEGF)Rpa}/J

Stock number: 033171
Product name: Agrp^DTR
Research areas: Neurobiology Research, Research Tools

Description

NPY/AGRP neurons regulate feeding behavior and body weight. Human diphtheria toxin receptor (HBEGF formerly DTR) and DsRed2 fluorescent reporter have been targeted to the mouse Agrp locus in these Agrp^DTR animals to create an Agrp knock-out allele and enable temporal ablation of AgP/NPY neurons after an injection of diptheria toxin.

Detailed description

Hypothalamic neurons that express neuropeptide Y (NPY) and agouti-related neuropeptide (AGRP) regulate feeding behavior and body weight.

Human diphtheria toxin receptor (HBEGF) and fluorescent DsRed2 reporter (detected with in situ hybridization) have been targeted to the mouse Agrp locus in these Agrp^DTR mice. This creates an Agrp knock-out allele and enables temporal control of NPY/AGRP neuron ablation after an injection of diptheria toxin. Homozygous Agrp^DTR/DTR mice show no obvious phenotype.

Treating either heterozygous or homozygous mice with diptheria toxin (DT) results in ablation of AGRP neurons. If this is done to neonatal mice, the mice survive. Treating adult mice with DT results in starvation within 6-8 days, however starvation can be prevented by a variety of manipulations, Wu et al (2012) PMID 22232663.

Development

A cassette containing the human HBEGF open reading frame followed by in internal ribosome entry site, a DsRed2 reporter gene, and loxP-flanked Sv-Neo (SV40 promoter-neomycin-phosphotransferase) gene was inserted just upstream of the initiation codon of Agrp in the first exon via homologous recombination in 129S4/SvJaeSor-derived AK18.1 embryonic stem (ES) cells. Resultant mice were bred with germline Meox2^tm1(cre)Sor (Stock No. 003755) mice to remove the Sv-Neo gene and create the Agrp^DTR allele. Meox2^tm1(cre)Sor was bred out of this line, and mice were backcrossed to C57BL/6J for more than 20 generations by the donating laboratory.

Allele

Symbol: Agrp^{tm2(HBEGF)Rpa}
Name: targeted mutation 2, Richard D Palmiter
MGI accession ID: MGI:3527177
Generation method: Targeted
Attributes: Reporter; Inserted expressed sequence
Synonyms: Agrp^DTR; Agrp^tm2(DTR)Rpa
Marker symbol: Agrp
Marker name: agouti related neuropeptide
Marker synonyms: agouti related protein; AGRT; ART; ASIP2
Chromosome: 8
Strain of origin: 129S4/SvJaeSor
Molecular note: The human diphtheria toxin receptor followed by an IRES-DsRed2 reporter gene was targeted to the Agrp locus to allow conditional killing of these cells. Germ-line, cre-mediated recombination was used to remove the neo cassette.