Coupling of vasopressin-induced intracellular Ca2+ mobilization and apical exocytosis in perfused rat kidney collecting duct

Arginine vasopressin (AVP) regulates the osmotic water permeability of the kidney collecting duct by inducing exocytotic insertion of aquaporin-2 into apical membrane. The coupling between AVP-induced intracellular Ca2+ mobilization and apical exocytosis was investigated in isolated perfused rat inner medullary collecting duct (IMCD) segments using confocal fluorescence microscopy. Changes of [Ca2+ ]i in IMCD cells were measured with fluo-4. A novel confocal imaging technique using a styryl dye, FM1-43, was developed to monitor real-time exocytosis induced by arginine vasopressin. AVP (0.1 nm) triggered a rapid increase of [Ca2+]i in IMCD cells, followed by sustained oscillations. Ratiometric measurement of [Ca2+]i confirmed that the observed [Ca2+]i oscillation was a primary event and was not secondary to changes in cell volume. The frequencies of [Ca2+]i oscillations in each IMCD cell were independent and time variant. 1-Deamino-8-d-arginine vasopressin (a V2 receptor agonist, 0.1 nm) simulated the effects of AVP by triggering [Ca2+]i oscillations. In the absence of extracellular Ca2+, ryanodine (0.1 mm) inhibited AVP-induced Ca2+ mobilization. AVP (0.1 nm) triggered accumulative apical exocytosis in IMCD cells within 20 s after application. Pre-incubating the IMCD with an intracellular Ca2+ chelator, BAPTA, prevented AVP-induced intracellular Ca2+ mobilization, apical exocytosis, and increase of osmotic water permeability. These results indicate that AVP, via the V2 receptor, triggers a calcium signalling cascade observed as [Ca2+]i oscillations in the IMCD and that intracellular Ca2+ mobilization is required for exocytotic insertion of aquaporin-2

Structural simplicity of the Zonula Occludens in the electrolyte secreting epithelium of the avian salt gland

The structure of the zonula occludens in the secretory epithelium of the salt gland of the domestic duck was determined by thin section and freeze-fracture electron microscopy. These glands secrete an effluent with a NaCl concentration four times that of plasma, and thus maintain a steep ionic gradient across their secretory epithelium. Freezefracture replicas from salt stressed ducks demonstrate that the zonula occludens is surprisingly shallow in depth (20–25 nm) and generally consists of two parallel junctional strands which are juxaposed along their entire length. In addition to the simplicity of the junction separating mucosal and serosal compartments, the ratio of junctional length to apical surface area is large since luminal surfaces of secretory cells are narrow and intermesh with one another. The zonula occludens in nonsecreting fresh water-adapted birds is similar to the salt stressed group except that two sets of double strand junctions are seen in addition to junctions consisting of a single set. Based on previous ultrastructural, cytochemical and physiological studies in salt glands and in other epithelia, a model for salt secretion was suggested in which intercellular space Na + , generated by basolateral ouabain-sensitive Na + pumps, reaches the lumen via a paracellular route (Ernst & Mills, 1977, J. Cell Biol. 75 :74). The simplicity of the morphological appearance of the zonula occludens in the salt gland, which resembles that described for several epithelia known to be leaky to ions, is consistent with this hypothesis.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/48028/1/232_2005_Article_BF01869292.pd