Ion channels from the midgut apical membrane of gypsy moth (Lymantria dispar) larvae were studied following mechanical fusion of brush-border membrane vesicles with planar phospholipid bilayer membranes. In symmetrical 300 mmol l(−)(1) KCl (pH 9.0), nine different channels with conductances ranging from 27 to 795 pS and linear current/voltage relationships were resolved. In the presence of a KCl gradient across the bilayer (450 mmol l(−)(1)cis/150 mmol l(−)(1)trans), 11 different conductance levels ranging from 16 to 850 pS were detected. The channels were slightly cationic: the zero-current reversal potential was shifted by −5 mV to −21 mV compared with symmetrical KCl conditions, corresponding to p(K)/p(Cl) permeability ratios of 1.5-8.0. Most channels were neither voltage-dependent nor Ca(2+)-sensitive and displayed complex gating kinetics. Addition of Ba(2+) or Cs(+) to both sides of the bilayer had little effect on channel activity, but fewer distinct channels were observed when KCl was replaced by potassium gluconate, suggesting an effect of Cl(−) on channel activity. A reduced number of channels was also detected when KCl was replaced by N-methyl-d-glucamine-HCl. Under asymmetrical N-methyl-d-glucamine-HCl conditions, only anionic channels were observed. They exhibited current rectification (35 pS at negative voltages and 81 pS at positive voltages) and were strongly voltage-dependent.
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