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| Item Type: | Article |
|---|---|
| Title: | Cell volume kinetics of adherent epithelial cells measured by laser scanning reflection microscopy: determination of water permeability changes of renal principal cells |
| Creators Name: | Maric, K., Wiesner, B., Lorenz, D., Klussmann, E., Betz, T. and Rosenthal, W. |
| Abstract: | The water channel aquaporin-2 (AQP2), a key component of the antidiuretic machinery in the kidney, is rapidly regulated by the antidiuretic hormone vasopressin. The hormone exerts its action by inducing a translocation of AQP2 from intracellular vesicles to the cell membrane. This step requires the elevation of intracellular cyclic AMP. We describe here a new method, laser scanning reflection microscopy (LSRM), suitable for determining cellular osmotic water permeability coefficient changes in primary cultured inner medullary collecting duct (IMCD) cells. The recording of vertical-reflection-mode x-z-scan section areas of unstained, living IMCD cells proved useful and valid for the investigation of osmotic water permeability changes. The time-dependent increases of reflection-mode x-z-scan section areas of swelling cells were fitted to a single-exponential equation. The analysis of the time constants of these processes indicates a twofold increase in osmotic water permeability of IMCD cells after treatment of the cells both with forskolin, a cyclic AMP-elevating agent, and with Clostridium difficile toxin B, an inhibitor of Rho proteins that leads to depolymerization of F-actin-containing stress fibers. This indicates that both agents lead to the functional insertion of AQP2 into the cell membrane. Thus, we have established a new functional assay for the study of the regulation of the water permeability at the cellular level. |
| Keywords: | Actins, Bacterial Proteins, Bacterial Toxins, Cell Line, Cyclic AMP, Forskolin, Kidney, Kinetics, Confocal Microscopy, Fluorescence Microscopy, Osmosis, Tertiary Protein Structure, Protein Transport, Time Factors, Water, Animals, Dogs, Rats |
| Source: | Biophysical Journal |
| ISSN: | 0006-3495 |
| Publisher: | Biophysical Society |
| Volume: | 80 |
| Number: | 4 |
| Page Range: | 1783-1790 |
| Date: | April 2001 |
| Official Publication: | https://doi.org/10.1016/S0006-3495(01)76148-6 |
| PubMed: | View item in PubMed |
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