Stimulatory effects of chlorzoxazone, a centrally acting muscle relaxant, on large conductance calcium-activated potassium channels in pituitary GH3 cells

Chlorzoxazone, a centrally acting muscle relaxant, has been used as a marker for hepatic CYP2E1 activity. However, little is known about the mechanism of chlorzoxazone actions on ion currents in neurons or neuroendocrine cells. We thus investigated its effects on ion currents in GH(3) lactotrophs. Chlorzoxazone reversibly increased Ca2+-activated K+ current (I-kappa(Ca)) in a concentration-dependent manner with an EC50 value of 30 muM. The chlorzoxazone-stimulated I-kappa(Ca) was inhibited by iberitoxin (200 nM) or clotrimazole (10 muM), but not by glibenclamide (10 muM) or apamin (200 nM). Chlorzoxazone (30 muM) suppressed voltage-dependent L-type Ca 21 current. In the inside-out configuration, chlorzoxazone applied to the intracellular side of the patch did not modify single-channel conductance of large conductance Ca2+-activated K+ (BKCa) channels, but did increase channel activity by increasing mean open time and decreasing mean closed time. Chlorzoxazone also caused a left shift in the activation curve of BKCa channels. However, Ca2+-sensitivity of these channels was unaffected by chlorzoxazone. 1-Ethyl-2-benzimidazolinone (30 muM), 2-amino-5-chlorobenzoxazole (30 muM) or chlormezanone (30 muM) enhanced BKCa channel activity, while 6-hydroxychlorzoxazone (30 muM) slightly increased it; however, chlorphenesin carbamate (30 muM) had no effect on it. Under the current-clamp condition, chlorzoxazone (10 muM) reduced the firing rate of action potentials. In neuroblastoma IMR-32 cells, chlorzoxazone (30 muM) also stimulated BKCa channel activity. The stimulatory effects of chlorzoxazone on these channels may be responsible for the underlying mechanism of chlorzoxazone actions on neurons and neuroendocrine cells. (C) 2002 Elsevier Science B.V. All rights reserved.