B6.129P2-P2ry2^tm1Bhk/J

Stock number: 009132
Product name: P2Y₂-R^-
Research areas: Cell Biology Research, Internal/Organ Research, Neurobiology Research, Research Tools, Sensorineural Research

Description

These purinergic receptor P2Y, G-protein coupled 2 (P2ry2^tm1Bhk) knock-out mice are susceptible to lung infections of Pseudomonas aeruginosa, exhibit impaired neutrophil chemotaxis, show a loss of sensitivity to thermal nociception and capsaicin, and have increased urinary concentration. This mutant mouse strain may be useful in studies of purinergic signaling in the regulation of ion transport, immune response to bacterial infections, pain response, neuronal differentiation and neurogenesis.

Detailed description

Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. No gene product (mRNA) is detected by Northern blot analysis of kidney tissue. Intracellular calcium ion levels in lung fibroblasts isolated from homozygotes fail to respond to nucleotide (UTP, ATP, ADP, UDP) challenge. Isolated airway epithelial cells have a loss of or diminished intracellular calcium response to nucleotide challenge. Unlike wild-type macrophages, macrophages derived from these animals fail to respond to extracellular stimulation with UTP. ATP- and adenosine 50[g-thio] triphosphate (ATPgS)-evoked aortic endothelium-dependent relaxation is reduced. Luminal nucleotide stimulated distal colonic ion transport is reduced. Homozygotes are more susceptible to lung infections of Pseudomonas aeruginosa, exhibit impaired neutrophil chemotaxis (loss in gradient sensing and migration distance), and loss of sensitivity to thermal nociception and capsaicin. Mutant mice fail to develop thermal hypersensitivity after to inflammatory injury (inflammatory hyperalgesia). Dorsal root ganglia exhibit diminished UTP-evoked Ca++ response. Growth-associated protein-43 (GAP-43) levels are not increased in sciatic nerves of mutants in response to ATPgammaS injection. Kidney function defects include increased renal collecting duct epithelial sodium channel activity and increased urine concentration. This mutant mouse strain may be useful in studies of purinergic signaling in the regulation of ion transport, immune response to bacterial infections, pain response, neuronal differentiation and neurogenesis.

Development

A targeting vector containing neomycin resistance and herpes simplex virus thymidine kinase genes was used to disrupt base pairs 552-1149 of the published cDNA sequence. The construct was electroporated into 129P2/OlaHsd derived E14TG2a embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The donating investigator reported that the resulting chimeric animals were crossed to C57BL/6NTac mice, and then backcrossed to the same for 12 generations (see SNP note below).

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 4 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.

Allele

Symbol: P2ry2^tm1Bhk
Name: targeted mutation 1, Beverly Koller
MGI accession ID: MGI:3613160
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: P2ry2
Marker name: purinergic receptor P2Y, G-protein coupled 2
Chromosome: 7
Strain of origin: 129P2/OlaHsd
Molecular note: A neomycin selection cassette replaced a genomic fragment corresponding to base pairs 552 - 1149 of the published cDNA. Northern blot analysis on RNA derived from kidney of homozygous mice confirmed that no detectable transcript is expressed from this allele.