NOD/ShiLt congenic mice carrying the Ncf1 spontaneous mutation are useful to study the role of free radicals, specifically superoxide production, on immune cells and autoimmune pathogenesis.
Dr. Clayton Mathews, University of Florida
On the C57BL/6 background, Peritoneal neutrophils and macrophages, bone marrow cells and neutrophils isolated from bone marrow of mice homozygous for Ncf1m1J fail to produce superoxide upon stimulation in vitro with N-formyl-methionyl-leucyl-phenylalanine (fMLP) or phorbol 12-myristate 13-acetate (PMA), as determined by kinetic spectrophotometric analysis of cytochrome c reduction. Western blot analysis detected no full-length NCF1/p47phox protein in cells from these mice. Analysis for other NADPH oxidases involved in neutrophil superoxide production revealed that NCF2/p67phox was present at wild type levels and CYBB/gp91phox and CYBA/p22phox were expressed at higher than wild type levels. Other neutrophil products were assayed and found not to differ in bone marrow cells of C57BL/6J vs. mutant mice; (Huang et al. 2000).
NOD.Ncf1 deficient mice lack functional NADPH oxidase enzymes and are unable to elicit respiratory burst. These mice exhibit reduced insulitis and reduced diabetes incidence. Mice carrying the Ncf1 spontaneous mutation are useful to study the role of free radicals, specifically superoxide production, on immune cells and autoimmune pathogenesis.
A Chr. 5 spontaneous mutation contains an A to C transversion at -2 position at the 5' end of exon 8 of the Ncf1 gene, resulting in aberrant splicing of the Ncf1 transcript. Immunoblotting detected no intact NCF1 protein in cells from these mice. The Ncf1 mutation arose in the B6.BKS(D)-Leprdb/J (Stock No. 000697) production colony at The Jackson Laboratory. Utilizing speed congenic technology, progeny of B6(Cg)-Ncf1m1J/J, Stock No. 004742, were crossed onto the NOD/ShiLt background. At N10 the mice were intercrossed to homozygosity. In 2010, the Type 1 Diabetes Resource received this strain at N10F6.
|Allele Name||mutation 1, Jackson|
|Allele Synonym(s)||Ncf1*; p47-; p47phox; p47phox-`; rs230824082|
|Gene Symbol and Name||Ncf1, neutrophil cytosolic factor 1|
|Strain of Origin||B6.Cg-Dock7m +/+ Leprdb/J|
|Molecular Note||The mutation is an A-to-C transversion at the -2 position 5' off exon 8 in intron 7, changing splice acceptor site CAG to CCG. This results in aberrant splicing of the transcript. Immunoblotting detected no intact NCF1 protein in cells from these mice.|
When using the NOD-Ncf1m1J mouse strain in a publication, please cite the originating article(s) and include JAX stock #010606 in your Materials and Methods section.