Measurements and modeling of water transport and osmoregulation in a single kidney cell using optical tweezers and videomicroscopy.
With an optical tweezer installed in our optical microscope we grab a single Madin Darby Canine kidney cell and keep it suspended in the medium without touching the glass substrate or other cells. Since the optically trapped cell remains with a closely round shape, we can directly measure its volume by using videomicroscopy with digital image analysis. We submit this cell to a hyperosmotic shock (up-shock) and video record the process: the cell initially shrinks due to osmotic efflux of water and after a while, due to regulatory volume increase (RVI), an osmoregulation response, it inflates again (water influx) until it reaches a new volume (the regulatory volume VR). In addition to considering standard osmotic water transport, we model RVI using a simple phenomenological model. We obtain an expression for cell volume variation as a function of time that fits very well with our experimental data, where two characteristic times appear naturally: one related to water transport and the other related to RVI. From the fit we obtain water permeability, osmolyte influx rate for RVI, and regulatory volume. With the addition of the hormone vasopressin, water permeability increases while the regulatory volume decreases until inhibition of RVI. In summary, we present a technique to measure directly volume changes of a single isolated kidney cell under osmotic shock and a phenomenological analysis of water transport that takes into account osmoregulation.
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Departamento de Física, ICEX, Universidade Federal de Minas Gerais, Caixa Postal 702, Belo Horizonte, CEP 30123-970 Minas Gerais, Brazil.
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D000222:Adaptation, Physiological / Q000187:drug effects / Q000502:physiology; D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology*; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003198:Computer Simulation; D004285:Dogs; D004867:Equipment Design; D005434:Flow Cytometry / Q000379:methods*; D007668:Kidney / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D007834:Lasers; D018715:Microscopy, Video / Q000379:methods; D008954:Models, Biological; D009997:Osmotic Pressure / Q000187:drug effects; D010812:Physical Stimulation / Q000295:instrumentation / Q000379:methods; D014667:Vasopressins / Q000494:pharmacology; D014867:Water / Q000378:metabolism*; D014882:Water-Electrolyte Balance / Q000187:drug effects / Q000502:physiology*
D014867:Water; D014667:Vasopressins