B6;129P2-Or8a1^{tm41(Drd1)Mom}/MomJ

Stock number: 027251
Research areas: Cell Biology Research, Developmental Biology Research, Neurobiology Research, Research Tools, Sensorineural Research

Description

This allele replaces the coding sequence of the G-protein coupled olfactory receptor Or8a1 (formerly called Olfr151) with the coding sequence of Drd1, a G-protein coupled receptor not usually expressed in the main olfactory epithelium. DRD1 shares 16% amino acid identity with OR8A1 and binds GNAS and GNAL. This substitution allele is useful for assessing the minimal sequence requirements for a G protein coupled receptor driven by the promoter of an olfactory sensory receptor to facilitate monoallelic expression of olfactory receptors, coalescence of olfactory glomeruli, and signal transduction via cAMP in olfactory sensory neurons.

Detailed description

This substitution allele is partially successful as a surrogate olfactory receptor, but not as good as the substitution of beta 2 adrenergic receptor (see Stock# 006734). Consistent with normal allelic exclusion of olfactory receptors, olfactory sensory neurons that express this GFP tagged MC4R substitution allele do not express any other olfactory receptor, proving this sequence adequate for monoallelic expression. However, the level of expression and number of olfactory neurons expressing this substitution allele are reduced compared with that of endogenous OR8A1 (formerly called OLFR151) on non-mutant olfactory sensory neurons, and immunohistological assessment found 58% of these GFP-marked neurons also expressed GMAP43 but not OMP indicative of an immature phenotype. Subcellular localization of this MC4R on olfactory sensory neurons is highly concentrated in the dendritic knob and endings of the cilia. Patch-clamp recordings show that these neurons respond to the MC4R selective agonist Ro27-3225. The axons from these neurons fasciculate but generally fail to coalesce into glomeruli.

Development

The coding sequence of Or8a1 (formerly called Olfr151) was replaced with the sequence encoding amino acides 1-446 of Drd1 and an IRES-tauGFP was inserted just downstream for bicistronic expression. The targeted mutation was generated in 129P2/OlaHsd-derived E14 ES cells. The ACNf neomycin cassette was self-excised by Cre recombination in the germline of the male chimeras, which were then bred with C57BL6/J females. After several generations of sibling breeding sperm was cryopreserved from homozygous males.

Allele

Symbol: Or8a1^{tm41(Drd1)Mom}
Name: targeted mutation 41, Peter Mombaerts
MGI accession ID: MGI:5616561
Generation method: Targeted
Attributes: Reporter; Null/Knockout; Inserted expressed sequence
Synonyms: Drd1a- M71-IRES-tauGFP
Marker symbol: Or8a1
Marker name: olfactory receptor family 8 subfamily A member 1
Marker synonyms: GA_x6K02T2PVTD-31408136-31407207; M71; MOR171-2; Olfr151; Olr1195; OR11-318; OST025
Chromosome: 9
Strain of origin: 129P2/OlaHsd
Expressed genes: GFP,Green Fluorescent Protein,; Drd1,dopamine receptor D1,mouse, laboratory
Site of expression: Olfactory sensory neurons.
Molecular note: The endogenous coding sequence was entirely replaced with the sequence encoding amino acids 1-446 of Drd1 and an IRES-tauGFP reporter cassette along with the self-excising ACNf selection cassette, so that this allele bicistronically expresses Drd1 and the tauGFP reporter.