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Modulation of neutrophil migration by exogenous gaseous nitric oxide
Author(s) -
VanUffelen Bart E.,
Koster Ben M.,
Broek Peter J. A.,
VanSteveninck John,
Elferink Jan G. R.
Publication year - 1996
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.60.1.94
Subject(s) - chemotaxis , extracellular , nitric oxide , intracellular , guanosine , cgmp dependent protein kinase , calcium , n formylmethionine leucyl phenylalanine , biology , biochemistry , calmodulin , calcium in biology , cyclic guanosine monophosphate , protein kinase a , biophysics , kinase , microbiology and biotechnology , endocrinology , medicine , receptor , enzyme , cyclin dependent kinase 2
We studied the effect of exogenous nitric oxide (NO) on migration of rabbit peritoneal neutrophils. Exogenous NO enhanced random migration of neutrophils in a concentration‐dependent way. An optimally stimulatory effect was observed with 0.5 μM NO, whereas at higher NO concentrations the enhancing effect decreased again. NO caused a rapid and transient increase in intracellular guanosine‐3′,5′‐cyclic monophosphate (cGMP) levels. The enhancing effect of NO on random migration was largely reversed by the inhibitors of cGMP accumulation, LY‐83583 and methylene blue, and by the antagonists of cGMP‐dependent protein kinase, 8‐bromoguanosine‐3′,5′‐cycHc monophosphorothioate, Rp‐isomer (Rp‐8‐Br‐cGMPS) and 8‐(4‐chlorophenylthio)‐guanosine‐3′,5′‐cyclic monophosphorothioate (Rp‐8‐pCPT‐cGMPS). These observations strongly suggest that the enhancement of random migration by NO is mediated by cGMP and cGMP‐dependent protein kinase. The effect of NO on migration did not occur in the absence of extracellular calcium. Although NO did not induce a measurable elevation of intracellular free calcium, preincubation with the intracellular calcium chelator Fura‐2/AM abolished the enhancing effect of NO. It appears therefore that a small change in the level of cytoplasmic free calcium does play a role in the enhancement of random migration by NO. High concentrations of NO were found to inhibit Chemotaxis induced by an optimal concentration of the chemotactic peptide AT‐formyl‐methionyl‐leucyl‐phenylalanine (fMLP). This inhibitory effect was also dependent on the presence of extracellular calcium. A role for cGMP in the inhibition of fMLP‐induced Chemotaxis by NO is not supported by our measurements of intracellular cGMP levels. In contrast to the effects on fMLP, NO did not affect Chemotaxis induced by the phorbol ester PMA. In conclusion, we show that NO, not derived from NO donors but applied directly, may stimulate or inhibit neutrophil migration, dependent on the concentration. The enhancing effect of NO on random migration is mediated by cGMP, which emphasizes the importance of this second messenger as a modulator of neutrophil functions.