Endotoxin-induced L-arginine pathway produces nitric oxide and modulates the Ca2+-activated K+ channel in cultured human dermal papilla cells.
Endotoxin includes an enzyme that synthesizes nitric oxide (NO) from l-arginine (NO synthase) in vascular smooth muscles cells, macrophages, and fibroblasts, leading to the release of NO. We evaluated the release of NO and its intracellular action on the Ca2+-activated K+ channel (KCa channel) in cultured human dermal papilla cells by use of the electron paramagnetic response (EPR) spin trapping method and the patch clamp technique. In dermal papilla cells pretreated for 24 h with endotoxin (1 microgram/microliter), application of 1 microM L-arginine generated NO, although no measurable release of NO was observed in cells without endotoxin pretreatment, as determined by the EPR spin trapping method. With the patch clamp technique, we found that the KCa channel of dermal papilla cells had high conductance and was voltage dependent. In addition, after endotoxin pretreatment, the extracellular application of 100 microM l-arginine modulated the KCa channel in the cell-attached patch configurations. In inside-out patch configuration, however, NO produced by L-arginine itself did not modulate the Kca channel. The modulation of the KCa channel was suppressed by pretreatment with 100 microM N omega-nitro-L arginine methyl ester, and inhibitor of inducible and constitutive NO synthases. Methylene blue, a blocker of guanylate cyclase, inhibited the L-arginine-induced activation of the Kca channel. Theses results indicate that the endotoxin-induced L-arginine pathway cell generates No, which consequently modulated the KCa channel in cultured human dermal papilla cells by increasing of cyclic GMP-dependent phosphorylation.
Nameda
Y
Y
Department of Dermatology, The University of Tokushima, Japan.
Miyoshi
H
H
Tsuchiya
K
K
Nakaya
Y
Y
Arase
S
S
D001120:Arginine / Q000031:analogs & derivatives / Q000187:drug effects / Q000494:pharmacology / Q000502:physiology*; D002118:Calcium / Q000494:pharmacology*; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000494:pharmacology; D004731:Endotoxins / Q000494:pharmacology*; D018859:Hair Follicle / Q000166:cytology; D006801:Humans; D019331:NG-Nitroarginine Methyl Ester; D009569:Nitric Oxide / Q000096:biosynthesis* / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology*; D011494:Protein Kinases / Q000187:drug effects; D012867:Skin / Q000166:cytology*
D004731:Endotoxins; D015221:Potassium Channels; D009569:Nitric Oxide; D001120:Arginine; D011494:Protein Kinases; D006152:Cyclic GMP; D002118:Calcium; D019331:NG-Nitroarginine Methyl Ester