Energizing Epithelial Transport with the Vacuolar H+-ATPase

The Ussing model has long provided the conceptual foundation for understanding epithelial transport mechanisms energized by the Na+-K+-ATPase. Plasma membranes may also use the vacuolar (V-type) H+-ATPase as the primary energy source of membrane and epithelial transport. A pure electrogenic pump, the V-type H+-ATPase energizes not only membranes it inhabits but also other transport pathways via electrical coupling.

Electrogenic pump and a Ca(2+)- dependent K+ conductance contribute to a posttetanic hyperpolarization in lamprey sensory neurons

1. Tetanic stimulation of lamprey sensory dorsal cells resulted in a posttetanic hyperpolarization (PTH). The amplitude and duration of the PTH were dependent on the stimulus duration and frequency. The PTH was not reversed at membrane potentials negative to -100 mV, whereas the afterhyperpolarization following single action potentials reversed at approximately -85 mV. There was also a biphasic effect on the input resistance during the PTH, with an early reduction that recovered to control before the PTH had decayed. 2. The amplitude and duration of the PTH were increased in Ringer solution containing tetraethylammonium and 4-aminopyridine, both of which broadened single action potentials, but were reduced after intracellular injection of Cs+. Ca(2+)-free Ringer solution, Cd2+, and Co2+ also reduced the PTH, suggesting the involvement of a Ca(2+)-dependent K+ conductance. However, the PTH was not reduced in Ba2+ Ringer solution, or by the Ca(2+)-dependent K+ channel antagonists apamin and charybdotoxin. 3. The cardiac glycoside ouabain reduced the amplitude and duration of the PTH, as did substitution of Na+ with choline or Li+. K(+)-free Ringer solution also reduced the PTH, whereas high-K+ Ringer solution had more variable effects. The amplitude and duration of the PTH were also dependent on temperature. These results support the involvement of an ouabain-sensitive Na-K pump in the PTH. 4. The PTH was reduced by the tachykinins substance P and physalaemin, and by 5-hydroxytryptamine, which blocks apamin-sensitive Ca(2+)-dependent K+ channels in the lamprey. However, gamma-aminobutyric acid, which has been reported to reduce a Ca(2+)-dependent K+ conductance in the dorsal cells, did not reduce the PTH. 5. These results suggest that a Ca(2+)-dependent K+ conductance and an Na-K electrogenic pump underlie the PTH. The PTH reduces the excitability of the dorsal cells, suggesting that it may act as a mechanism to gate sensory information entering the spinal cord.

Serotonin depresses the after-hyperpolarization through the inhibition of the Na+/K+ electrogenic pump in T sensory neurones of the leech

In T sensory neurones of the leech, a train of impulses elicited by intracellular electrical stimulation leads to an after-hyperpolarization of up to 30 mV, mainly due to the activation of the electrogenic Na+/K(+)-ATPase but partly to a Ca2(+)-activated K+ conductance. It was found that serotonin reversibly reduced the amplitude of this after-hyperpolarization. We investigated the mechanism of action of serotonin and found: (1) after inhibition of the Ca2(+)-activated K+ conductance with BaCl2 or CdCl2, serotonin was still able to reduce the after-hyperpolarization; (2) when penetration of T cells with microelectrodes leaking sodium was preceded by serotonin perfusion of the ganglia, the normal hyperpolarization due to the activation of the electrogenic pump was converted to a depolarization; (3) after long-lasting perfusion with K(+)-free saline solution (which inhibits the Na+/K+ pump), the application of CsCl caused repolarization by reactivating the electrogenic ATPase; serotonin slowed and reduced this repolarization; (4) serotonin potentiated the depolarization of T neurones caused by the inhibition of the Na+/K+ pump following cooling of ganglia and depressed the hyperpolarization after rewarming to room temperature. These data taken together suggest that serotonin directly inhibits the Na+/K+ electrogenic pump.