1. From two lines of evidence, we conclude that the potassium transport gives rise directly to the midgut potential, i.e. that the active potassium transport mechanism is electrogenic. 2. First, diffusion potentials of neither potassium, sodium, magnesium, calcium, nor chloride could give rise to the large midgut potential if values for tissue concentrations are accepted for their respective activities in the epithelium. 3. Secondly, no externally added cation other than potassium is required to sustain either the potential or short circuit current, no specific anion is required, and no metabolic ion is known to be produced in sufficient amount to act as a counter ion for potassium in a non-electrogenic process. 4. Changes in the concentration of potassium on the blood-side of the midgut always lead to changes in potential in the direction predicted by the Nernst equation. Moreover, a tenfold change in potassium concentration leads to the expected 59 mV. potential change provided that the prior midgut potential is at least 130 mV. This effect could be attributed either to the stimulation of an electrogenic potassium pump or to a potassium diffusion potential across the blood-side barrier.
1. The electrical potential across the isolated midgut of five developmental stages of the Cecropia silkworm was studied by changing the concentration of single cations in solutions bathing each side of the midgut. The stages included feeding fourth-instar insects, insects moulting from the fourth to the fifth instar, feeding fifth-instar insects, insects which had evacuated their midguts, and insects spinning cocoons. 2. Average values of the initial maximal potential exhibited by the midgut in solutions containing K, Mg, and Ca but no Na, for the stages mentioned above, were 68, 83, 90, 124, and 2 mV., respectively. 3. In all of the developmental stages studied except the ‘spinning larva’, reducing the potassium concentration from 32 to 2 mM/l. on the blood-side of the isolated gut lowers the potential, on the lumen-side of the gut raises the potential and on both sides gives an intermediate value. 4. When the potential prior to a decrease in concentration of potassium on the blood-side is over 100 mV., the Nernst slope approaches 59 mV. 5. A tenfold reduction in the concentration of magnesium or the addition of 32 mM/l. sodium to the solutions bathing the isolated gut has no systematic effect on the potential. 6. A tenfold drop in the concentration of calcium in the solutions causes changes in the potential in the opposite direction from those predicted by the Nernst equation. 7. The pH of the midgut contents rises from early fourth instar to late fifth instar. The hydrogen-ion concentration of the blood is about 1000 times more than that of midgut contents in fifth-instar insects. 8. Neither synthetic ecdysone, partially purified natural ecdysone nor juvenile hormone has an effect on the potential or current of the isolated midgut over periods as long as 30 min.
1. Flux measurements with 42 K reveal that in the isolated midgut of Hyalophora cecropia 90 to 100 % of the short-circuit current is carried by the active transport of potassium from the blood-side to the lumen. 2. When K-transport is strongly depressed, either by withholding potassium from the blood side or by imposing a large positive potential on the lumen, the oxygen uptake of the isolated gut remains virtually unchanged. If the K-transport were to be energized by the negligible increase in oxygen uptake about 40 µ -equiv. of potassium would have to be transported for every µ -equiv. of extra oxygen taken up. This ratio of K-transport to oxygen uptake is thermodynamically impossible. 3. The ratio of potassium transported to total oxygen consumed when the midgut is bathed with 32 mM potassium on both sides is about 1.3 at temperatures of 25° and 15° C. The ratio must be smaller at lower potassium concentrations and is 2.0 at 73.5 mM-K, which may be approaching the maximum value. 4. Although the oxygen uptake is independent of the K-transport, the reverse is not true. There is a close dependency of K-transport on oxygen consumption. 5. K-transport by the midgut contrasts with Na-transport by the frog skin because Na-transport stimulates oxidative metabolism whereas K-transport does not. Evidently the coupling of transport to energy supply is different in the two systems.