K+ CHANNEL PERMEATION AND BLOCK IN THE MIDGUT EPITHELIUM OF THE TOBACCO HORNWORM MANDUCA SEXTA

The K+-secreting larval midgut of Manduca sexta in vitro was voltage- or current-clamped. In contrast to Tl+, NH4+ and Na+, both Rb+ and K+ generated a short-circuit current, although with different saturation kinetics. The dependence of the short-circuit current on Rb+/K+ mole fraction gave no evidence for multi-ion occupation of the basolateral K+ channels. After 'functionally' eliminating the apical membranes using the ionophore amphotericin B and the 'apical K+ pump' blockers trimethyltin chloride or Tl+, the K+ channels could be more closely investigated. By measuring zero-current potentials, permeability ratios PX/PK were estimated using an adapted version of the Goldman­Hodgkin­Katz voltage equation. Their sequence was K+ (1) = Tl+ > Rb+ (0.38) > NH4+ (~0.3) > Cs+ (0.03) > Na+ (~0). The K+ channels could not be blocked by basally applied Cs+, Na+ or tetraethylammonium. Blockade of K+ current by Ba2+ was typically voltage-dependent, but only at moderate transbasal voltages. The relative electrical distance delta of the Ba2+ binding site from the basal channel opening was determined to be 0.2. At zero transbasal voltage, the apparent inhibition constant for barium KBa* was 1.7 mmol l-1.