Effects of potassium-free media on ADH action in toad urinary bladder

We studied the effects of potassium-free media on processes related to the hydro-osmotic response of toad bladder to ADH (20 mU/ml). Exposure of bladders to potassium-free media did not affect base-line osmotic water flow, but it promptly attenuated the level of osmotic water permeability induced by ADH. Both the frequency of hormonally induced intra(luminal)membrane particle aggregates (presumed sites for transmembrane water flow) and the number of luminal membrane fusion events (associated with aggregate delivery from the cytoplasm) were also reduced. Potassium-free media had no measurable effect either on cytoplasmic microtubule integrity or on mean aggregate size. Potassium repletion reversed the inhibitory effect of potassium-free media on ADH-related osmotic water permeability. For bladders fully stimulated with ADH in the presence of potassium, subsequent bathing media depletion of potassium led to an inhibition of ADH-related water flow and to reductions in membrane fusion sites and aggregates. We confirmed that the inhibitory effect of potassium-free media on ADH-induced osmotic water permeability results from serosal bathing medium potassium depletion alone and occurs at a post-cyclic AMP site. In addition, we found that ADH-stimulated water permeability was attenuated in bathing media containing a low potassium concentration (0.5 mM). The data are consistent with the view that potassium-free media or media containing low levels of potassium inhibit ADH-enhanced osmotic water permeability in toad bladder by interfering with the process of or leading to membrane fusion required for the delivery of water-conducting structures to the luminal membrane. In addition, some of our results imply that aggregates may turn over during sustained ADH stimulation.