BKS.129P2(Cg)-Nos3^tm1Unc/J

Stock number: 018295
Product name: eNOS-
Research areas: Cardiovascular Research, Diabetes and Obesity Research, Endocrine Deficiency Research, Internal/Organ Research, Metabolism Research, Mouse/Human Gene Homologs, Research Tools

Description

These C57BLKS.eNOS^- mice are a C57BLKS-congenic strain with a knockout mutation of the nitric oxide synthase 3 endothelial cell gene (Nos3 or eNOS). They may be useful in studying albuminuria/diabetic nephropathy, insulin resistance, hyperlipidemia, hypertension and other cardiovascular defects, wound healing, and lung development.

Detailed description

These C57BLKS/J-congenic mice harbor a knockout mutation of the Nos3 gene. Homozygous mice (C57BLKS.eNOS^-/-) are viable and fertile. C57BLKS.eNOS^-/- mice develop moderate albuminuria (by 26 weeks of age) and moderate systemic hypertension (by 24-26 weeks of age).

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. This is the case for the C57BLKS.eNOS^-/- strain. It should be noted that the C57BLKS.eNOS^-/- phenotype could vary from that originally described for eNOS^-/- mice on a C57BL/6-congenic background (described below). We may modify the C57BLKS.eNOS^-/- strain description if necessary as published results become available.

When maintained on a C57BL/6-congenic background, eNOS^-/- mice exhibit elevated blood pressure that is about 20 mmHg higher than that seen in normal wildtype siblings. They also show a decreased heart rate. Female homozygotes are smaller in body weight than normal wildtype siblings. Hyperglycemic-euglycemic clamp studies demonstrate that homozygotes exhibit insulin resistance at the level of the liver and peripheral tissues.

Development

These C57BLKS.eNOS^- mutant mice were generated by breeding males from the eNOS^- C57BLKS/J Lepr^db double mutant colony (Stock No. 008340) with C57BLKS/J inbred females (Stock No. 000662). The resulting offspring retaining the eNOS^- mutation and wildtype at the Lepr locus were bred together to establish this C57BLKS.eNOS^- colony as Stock No. 018295.

The eNOS knockout mutation was created by Dr. Oliver Smithies (University of North Carolina) as described for Stock No. 002684). It has a neomycin resistance cassette replacing 129 bp of exon 12 - this disrupted the calmodulin binding site of the protein and introduced a premature stop codon into the transcripts.

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. One marker on Chromosome 5 was still segregating with the targeted mutation. Two markers on Chromosome 11 were found to still be segregating with C57BL/6J as in parental line Stock No. 008340.

Allele

Symbol: Nos3^tm1Unc
Name: targeted mutation 1, University of North Carolina
MGI accession ID: MGI:1857229
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Nos3
Marker name: nitric oxide synthase 3, endothelial cell
Chromosome: 5
Strain of origin: 129P2/OlaHsd
Molecular note: A 1.2 kb neomycin cassette replaced 129 bp of exon 12 of the gene. This disrupted the calmodulin binding site of the protein and introduced a premature stop codon into the transcripts. Immunohistochemisty of heart and kidney sections from homozygous mutant mice confirmed that no detectable encoded protein was present.