Oxygen radical generation and expression of NADPH oxidase genes in bottlenose dolphin (Tursiops truncatus) neutrophils.
Oxygen radical generation by stimulation with phorbol myristate acetate (PMA) was evaluated in bottlenose dolphin neutrophils. A Cypridina luciferin analog-dependent chemiluminescent assay demonstrated that dolphin neutrophils generate superoxide by the addition of PMA, and that its superoxide-forming activity is completely suppressed by diphenylene iodonium, a specific inhibitor of NADPH oxidase. These results indicate that dolphin neutrophils possess NADPH oxidase activity. Furthermore, the NADPH oxidase activity (hydrogen peroxide production) in dolphin neutrophils, as well as in human neutrophils, was greater at 37 degrees C than at a lower temperature. RT-PCR with specific primers revealed that dolphin neutrophils expressed the mRNAs of the major NADPH oxidase components, which included membrane-associated flavocytochrome b (gp91(phox) and p22(phox)) and cytosolic factors (p40(phox), p47(phox), and p67(phox)), implying the existence of these protein homologues in dolphin neutrophils.
Itou
T
T
Department of Preventive Veterinary Medicine and Animal Health, Nihon University School of Veterinary Medicine, Nihon University, 1866 Kameino, Kanagawa 252-8510, Fujisawa, Japan. takuya@brs.nihon-u.ac.jp
Sugisawa
H
H
Inoue
Y
Y
Jinbo
T
T
Sakai
T
T
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D004289:Dolphins / Q000097:blood / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005609:Free Radicals / Q000097:blood; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D019255:NADPH Oxidases / Q000235:genetics*; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology*; D009861:Onium Compounds / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D013481:Superoxides / Q000097:blood*; D013696:Temperature; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D004791:Enzyme Inhibitors; D005609:Free Radicals; D009861:Onium Compounds; D012333:RNA, Messenger; D013481:Superoxides; C007517:diphenyleneiodonium; D006861:Hydrogen Peroxide; D019255:NADPH Oxidases; D013755:Tetradecanoylphorbol Acetate