Nitric oxide production by nurse shark (Ginglymostoma cirratum) and clearnose skate (Raja eglanteria) peripheral blood leucocytes.
Reactive nitrogen intermediates, such as nitric oxide (NO), are important immunomodulators in vertebrate immune systems, but have yet to be identified as mediators of host defence in any member of class Chondrichthyes, the cartilaginous fishes. In the present study, production of NO by nurse shark (Ginglymostoma cirratum) peripheral blood leucocytes (PBL) stimulated with bacterial cell wall lipopolysaccharide (LPS) was investigated. PBL were cultured for 24 to 96 h following stimulation with LPS at concentrations ranging from 0 to 25 microg ml(-1), in both serum-supplemented and serum-free culture conditions. Production of NO was measured indirectly using the Griess reaction, with maximal NO production occurring after 72 h using 10% FBS and 10 microg LPS ml(-1). Application of these culture conditions to PBL from another cartilaginous fish (clearnose skate, Raja eglanteria) resulted in a similar NO response. Addition of a specific inhibitor of inducible nitric oxide synthase (iNOS), L-N(6)-(1-iminoethyl)lysine (L-NIL), resulted in a significant decrease in the production of NO by PBL from both species.
Walsh
Cathy J
CJ
Mote Marine Laboratory, Sarasota, FL 34236, USA. cjwalsh@mote.org
Toranto
Jason D
JD
Gilliland
C Taylor
CT
Noyes
David R
DR
Bodine
Ashby B
AB
Luer
Carl A
CA
D000704:Analysis of Variance; D000818:Animals; D003124:Colorimetry; D005431:Florida; D007962:Leukocytes / Q000378:metabolism*; D008070:Lipopolysaccharides; D008239:Lysine / Q000031:analogs & derivatives / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors; D009573:Nitrites / Q000378:metabolism; D012754:Sharks / Q000378:metabolism*; D012860:Skates, Fish / Q000378:metabolism*
D008070:Lipopolysaccharides; C090663:N(6)-(1-iminoethyl)lysine; D009573:Nitrites; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D008239:Lysine