Mercurial reagents inhibit flow through ADH-induced water channels in toad bladder.

Mercurial reagents inhibit the water permeability of erythrocytes and proximal renal tubule. We examined the effect of two such agents on vasopressin-induced water transport across toad urinary bladder. Water flows were measured in unfixed tissues and in tissues fixed either with N-ethylmaleimide (NEM) or with glutaraldehyde. When added concurrently with 20 mU/ml vasopressin, 1 mM mucosal p-chloromercuribenzene-sulfonic acid (p-CMBS) inhibited water flow within 1 h. p-CMBS also inhibited flow in tissues that had been fixed with mucosal NEM after stimulation with vasopressin. However, p-CMBS did not affect flow in glutaraldehyde-fixed tissues. In contrast, HgCl2 inhibited water flow and urea permeability even in tissues that had been fixed with glutaraldehyde after stimulation with vasopressin. Inhibition was more pronounced when HgCl2 was added to the mucosal rather than the serosal bathing medium and was not reversed by dithiothreitol. HgCl2 did not diminish the frequency or area of luminal membrane aggregates observed by freeze-fracture electron microscopy. HgCl2 also did not affect amphotericin-induced water permeability in glutaraldehyde-treated tissues, suggesting that it did not diminish the permeability of cellular barriers to flow. Our results parallel closely those reported by other investigators for water flow across erythrocytes and proximal renal tubule and suggest that mercurial reagents can directly block the vasopressin-induced water channel. The water channel at the apical membrane of the toad bladder may prove to share structural similarity with that constantly present in erythrocytes and proximal renal tubule.