title: Amiloride-sensitive trypsinization of apical sodium channels. Analysis of hormonal regulation of sodium transport in toad bladder.
abstract: Incubation of the mucosal surface of the toad urinary bladder with trypsin (1 mg/ml) irreversibly decreased the short-circuit current to 50% of the initial value. This decrease was accompanied by a proportionate decrease in apical Na permeability, estimated from the change in amiloride-sensitive resistance in depolarized preparations. In contrast, the paracellular resistance was unaffected by trypsinization. Amiloride, a specific blocker of the apical Na channels, prevented inactivation by trypsin. Inhibition of Na transport by substitution of mucosal Na, however, had no effect on the response to trypsin. Trypsinization of the apical membrane was also used to study regulation of Na transport by anti-diuretic hormone (ADH) and aldosterone. Prior exposure of the apical surface to trypsin did not reduce the response to ADH, which indicates that the ADH-induced Na channels were inaccessible to trypsin before addition of the hormone. On the other hand, stimulation of short-circuit current by aldosterone or pyruvate (added to substrate-depleted, aldosterone-repleted bladders) was substantially reduced by prior trypsinization of the apical surface. Thus, the increase in apical Na permeability elicited by aldosterone or substrate involves activation of Na channels that are continuously present in the apical membrane in nonconductive but trypsin-sensitive forms.
mesh_terms: D000450:Aldosterone / Q000494:pharmacology*; D000584:Amiloride / Q000494:pharmacology*; D000818:Animals; D001692:Biological Transport; D002024:Bufo marinus / Q000502:physiology*; D007473:Ion Channels / Q000187:drug effects*; D011719:Pyrazines / Q000494:pharmacology*; D011773:Pyruvates / Q000494:pharmacology; D013268:Stimulation, Chemical; D014357:Trypsin / Q000378:metabolism* / Q000494:pharmacology; D001743:Urinary Bladder / Q000187:drug effects / Q000378:metabolism*; D014667:Vasopressins / Q000494:pharmacology*
chemical_list: D007473:Ion Channels; D011719:Pyrazines; D011773:Pyruvates; D014667:Vasopressins; D000450:Aldosterone; D000584:Amiloride; D014357:Trypsin