Effect of hypertonicity and colchicine on intramembranous particle aggregation in toad urinary bladder.

Abstract

Colchicine, an agent which disrupts microtubules, inhibits the vasopressin (VP)-induced increase in water permeability as well as intramembranous particle (IMP) aggregation in the luminal plasma membrane of granular cells of toad urinary bladder. However, the hydroosmotic response induced by serosal hypertonicity is not affected by colchicine. The present investigation was initiated to establish whether serosal hypertonicity is associated with IMP aggregation and whether the aggregation, if present, is altered by colchicine. The experimental half of paired hemibladders from the toad, Bufo marinus, treated with 0.1 mM colchicine for 4 h prior to exposure to serosal mannitol (240 mM) demonstrated no significant difference in osmotic water How (Jv) (1.03 × 0.18 vs. 1.13 ± 0.22μl · min−1 · cm−2; p>0.20) when compared with control hemibladders. Similarly, comparison of control and colchicine-treated bladders revealed no difference in the number of IMP aggregation sites per area of membrane (17.8 ± 2.0 vs. 24.7 ± 3.5/100μm; p>0.10), the relative area of membrane occupied by these sites (0.30 ± 0.06 vs. 0.39 ± 0.07%; p>0.10) or the mean size of the aggregates (17.0 ± 1.4 vs. 15.8 ± 1.0 × 103 μm2; p > 0.20). These results indicate that in toad bladder the increase in Jv induced by serosal hypertonicity is associated with IMP aggregation. Secondly, an intact microtubule system is not required to induce the hydroosmotic or the aggregation responses. If, as has been proposed, the cellular actions of VP and serosal hypertonicity share a common pathway to bring about an increase in osmotic water permeability and cause IMP aggregation in the luminal membrane of the granular cell, the present results suggest that the pathway begins at a step subsequent not only to the generation of cAMP, but also beyond the involvement of the microtubule system.