Membrane traffic and sorbitol release during osmo- and volume regulation in isolated rat renal inner medullary collecting duct cells.
In response to hypotonic stress, cells of the inner medullary collecting duct (IMCD) undergo swelling followed by a regulatory volume decrease (RVD) and a transient release of organic osmolytes such as sorbitol. In this study, we tested the hypothesis whether membrane recycling is involved in the latter process. Therefore, the state of submembranal actin and the cellular uptake or release of the fluid-phase marker fluorescein isothiocyanate (FITC)-dextran (FD) were investigated as related to changes in membrane permeability for sorbitol. After exposure to hypotonic medium the submembranal actin web rapidly disaggregated but it started to reorganize after 5 min of incubation. The basal-lateral pole of IMCD cells showed a significant uptake of extracellular FD after 100 sec. After 5 min, part of this fluorescence intensity had moved towards the cell center but the main part remained submembranal. Disintegration of the actin network by cytochalasin D diminished the uptake of FD during hypotonicity as did a permanent increase in intracellular calcium induced by ionomycin treatment. During a second osmotic stimulation of IMCD cells preloaded with FD, FD was released in a linear time course reaching a plateau after 1 min. Isotonic ionomycin treatment of preloaded cells also generated a rapid FD release during the first minute but induced a further 2-fold increase during the next 4 min. Under both conditions initial FD release was highly correlated with the simultaneously determined increase in sorbitol permeability. A similar strong correlation was found when different incubation temperatures were used (0 degree C, 15 degrees C, 37 degrees C). These results suggest that during exposure of IMCD cells to hypotonicity the submembranal actin web rapidly disintegrates, and "reserve" vesicles, probably containing sorbitol transporter, move to and fuse with the basal-lateral plasma membrane. The fusion causes a rapid increase in sorbitol permeability. These membrane areas are recovered by internalization, and the transport systems for sorbitol are concomitantly retrieved. In parallel to this internalization the submembranal actin filament network is rearranged. This process seems to be regulated by changes in intracellular calcium.
Czekay
R P
RP
Max-Planck-Institut für molekulare Physiologie, Dortmund/Germany.
Kinne-Saffran
E
E
Kinne
R K
RK
D000199:Actins / Q000032:analysis / Q000378:metabolism / Q000648:ultrastructure; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002463:Cell Membrane Permeability / Q000502:physiology; D015638:Cytochalasin D / Q000494:pharmacology; D003911:Dextrans / Q000493:pharmacokinetics; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D007038:Hypotonic Solutions / Q000494:pharmacology; D015759:Ionomycin / Q000494:pharmacology; D007685:Kidney Tubules, Collecting / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D008297:Male; D008870:Microtubules / Q000502:physiology / Q000648:ultrastructure; D009994:Osmolar Concentration; D051381:Rats; D017208:Rats, Wistar; D013012:Sorbitol / Q000378:metabolism* / Q000493:pharmacokinetics; D013696:Temperature
D000199:Actins; D003911:Dextrans; D007038:Hypotonic Solutions; C015219:fluorescein isothiocyanate dextran; D015638:Cytochalasin D; D013012:Sorbitol; D015759:Ionomycin; D016650:Fluorescein-5-isothiocyanate; D002118:Calcium