title: P47(phox)-deficient NADPH oxidase defect in neutrophils of diabetic mouse strains, C57BL/6J-m db/db and db/+.
abstract: Deficiencies in neutrophil NADPH oxidase proteins have been demonstrated in humans with chronic granulomatous disease. However, no spontaneous mutation in murine NADPH oxidase has been reported. In this study we report that neutrophils from the diabetic mouse strains, C57BL/6J-m heterozygous lean (lepr(db/+)) and homozygous obese (lepr(db/db)) mice produced no superoxide on stimulation. An absence of intact p47(phox) but not other oxidase proteins was observed in both mouse strains through the use of immunoblotting. Molecular analysis by reverse transcriptase-polymerase chain reaction identified three abnormal p47phox mRNA transcripts. Sequencing of genomic DNA of p47(phox) revealed a point mutation at the -2 position of exon 8, which is consistent with aberrant splicing of the p47(phox) transcript. These results indicate that the C57BL/6J-m db/db and db/+ mice are the first spontaneously derived murine model of NADPH oxidase deficiency involving a p47(phox) mutation.
authors.affiliation: Department of Pathology, University of Connecticut Health Center, Farmington 06030-3105, USA.
authors.forename: C K
authors.initials: CK
authors.lastname: Huang
authors.forename: L
authors.initials: L
authors.lastname: Zhan
authors.forename: M O
authors.initials: MO
authors.lastname: Hannigan
authors.forename: Y
authors.initials: Y
authors.lastname: Ai
authors.forename: T L
authors.initials: TL
authors.lastname: Leto
mesh_terms: D000818:Animals; D001483:Base Sequence; D003920:Diabetes Mellitus / Q000201:enzymology* / Q000235:genetics; D005260:Female; D005838:Genotype; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D019255:NADPH Oxidases / Q000737:chemistry / Q000172:deficiency*; D009504:Neutrophils / Q000201:enzymology*; D009765:Obesity / Q000201:enzymology / Q000235:genetics; D010750:Phosphoproteins / Q000172:deficiency* / Q000235:genetics; D017354:Point Mutation; D012326:RNA Splicing; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013481:Superoxides / Q000378:metabolism
chemical_list: D010750:Phosphoproteins; D012333:RNA, Messenger; D013481:Superoxides; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1