Yc. Kim et al., EFFECTS OF MYOSIN LIGHT-CHAIN KINASE INHIBITORS ON CARBACHOL-ACTIVATED NONSELECTIVE CATIONIC CURRENT IN GUINEA-PIG GASTRIC MYOCYTES, Pflugers Archiv, 434(4), 1997, pp. 346-353
The effects of myosin light chain kinase inhibitors on muscarinic stim
ulation-activated nonselective cationic current (I-CCh) in guinea-pig
gastric antral myocytes were studied using the whole-cell patch-clamp
technique. I-CCh was induced by carbachol (CCh; 50 mu M) at a holding
potential of -30 mV or -60 mV, ML-7, a chemical inhibitor of myosin li
ght chain kinase (MLCK), inhibited I-CCh concentration dependently in
a reversible manner (53 +/- 8.6% at 1 mu M, mean +/- SE, n = 11). in a
ddition, amplitudes of I-CCh were only 37 +/- 2.7% of the daily contro
l values following the addition of a peptide inhibitor of MLCK to the
pipette solution. On the other hand, ML-7 had an inhibitory effect on
voltage-operated Ca2+ channel current. The peak value of Ba2+ current
at 0 mV was reduced to 35 +/- 7.4% (n = 9) by 3 mu M of ML-7. As I-CCh
is known to have an intracellular Ca2+ dependence, we tried to exclud
e the possibility that ML-7 inhibited I-CCh, indirectly via suppressio
n of Ca2+ current and the similar inhibitory effects of ML-7 on I-CCh
were confirmed under the following conditions: (1) clamp of membrane p
otential at -60 mV; (2) clamp of intracellular [Ca2+] to 1 mu M by 10
mM BAPTA; (3) pre-inhibition of Ca2+ channel by verapamil. Different f
rom the effects on I-CCh ML-7 barely inhibited the same cationic curre
nt induced by guanosine 5'-O-(3-thiotriphosptlate) (GTP[gamma S], 0.2
mM) in the pipette solution. These results suggest that a Ca2+/calmodu
lin-MLCK-dependent pathway can modulate the activation of I-CCh in gui
nea-pig gastric antral myocytes.