Charybdotoxin block of Ca(2+)-activated K+ channels in colonic muscle depends on membrane potential dynamics.
Charybdotoxin (ChTX) is a specific blocker of Ca(2+)-activated K+ channels. The voltage- and time-dependent dynamics of ChTX block were investigated using canine colonic myocytes and the whole cell patch-clamp technique with step and ramp depolarization protocols. During prolonged step depolarizations, K+ current slowly increased in the continued presence of ChTX (100 nM). The rate of increase depended on membrane potential with an e-fold change for every 60 mV. During ramp depolarizations, the effectiveness of ChTX block depended significantly on the rate of the ramp (50% at 0.01 V/s to 80% at 0.5 V/s). Results are consistent with a mechanism in which ChTX slowly "unbinds" in a voltage-dependent manner. A simple kinetic model was developed in which ChTX binds to both open and closed states. Slow unbinding is consistent with ChTX having little effect on electrical slow waves recorded from circular muscle while causing depolarization and contraction of longitudinal muscle, which displays more rapid "spikes." Resting membrane potential and membrane potential dynamics are important determinants of ChTX action.
Frey
B W
BW
Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno 89557, USA.
Carl
A
A
Publicover
N G
NG
D000818:Animals; D018999:Charybdotoxin / Q000494:pharmacology*; D003106:Colon / Q000187:drug effects* / Q000378:metabolism; D004285:Dogs; D005260:Female; D066298:In Vitro Techniques; D051036:Large-Conductance Calcium-Activated Potassium Channels; D008297:Male; D008564:Membrane Potentials; D008954:Models, Biological; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015221:Potassium Channels / Q000187:drug effects* / Q000378:metabolism; D024681:Potassium Channels, Calcium-Activated
D051036:Large-Conductance Calcium-Activated Potassium Channels; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D018999:Charybdotoxin