Journal
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY
Volume 144, Issue 1, Pages 11-17Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.cbpb.2006.01.007
Keywords
bactericidal activity; nitric oxide; nitric oxide synthase; hemocyte; lipopolysaccharide; crustacean
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We determined the biochemical characteristics of nitric oxide synthase (NOS) in hemocytes of the crayfish Procambarus clarkii and investigated the roles of hemocyte-derived NO in host defense. Biochemical analysis indicated the presence of a Ca2+ -independent NOS activity, which was elevated by lipopolysaccharide (LPS) treatment. When bacteria (Staphylococcus aureus) and hemocytes were co-incubated, adhesion of bacteria to hemocytes was observed. NO donor sodium nitroprusside (SNP) significantly increased the numbers of hemocytes to which bacteria adhered. Similarly, LPS elicited bacterial adhesion and the LPS-induced adhesion was prevented by NOS inhibitor N-G-monomethyl-L-arginine (L-NMMA). Finally, plate count assay demonstrated that addition of LPS to the hemocytes/bacteria co-incubation resulted in a significant decrease in bacterial colony forming unit (CFU), and that L-NMMA reversed the decreasing effect of LPS on CFU. The combined results demonstrate the presence of a Ca2+-independent LPS-inducible NOS activity in crayfish hemocytes and suggest that hemocyte-detived NO is involved in promoting bacterial adhesion to hemocytes and enhancing bactericidal activity of hemocytes. (c) 2006 Elsevier Inc. All rights reserved.
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