Diffusive water permeability in isolated kidney proximal tubular cells: nature of the cellular water pathways.
The diffusive water permeability (Pd) of the plasma membrane of proximal kidney tubule cells was measured using a 1H-NMR technique. The values obtained for the exchange time (Tex) across the membrane were independent of the cytocrit and of the Mn2+ concentration (in the range 2.5 to 5 mM). At 25 degrees C the calculated Pd value was (per cm2 of outer surface area without taking into account membrane invaginations) 197 +/- 17 microns/sec. This value equals 22.3 +/- 1.9 microns/sec when the invaginations are taken into account. Cell exposure to 2.5 mM parachloromercuribenzenesulfonic acid, pCMBS, (for 20 to 35 min) reduced Pd to 45% of its control value. Five mM dithiothreitol, DTT, reverted this effect. The activation energy for the diffusive water flux was 5.2 +/- 1.0 kcal/mol under control conditions. It increased to 9.1 +/- 2.2 kcal/mol in the presence of 2.5 mM pCMBS. Using our previous values for the osmotic water permeability (Pos) in proximal straight tubular cells the Pos/Pd ratio equals 18 +/- 1, under control conditions, and 3.2 +/- 0.3 in the presence of pCMBS. These experimental results indicate the presence of pathways for water, formed by proteins, crossing these membranes, which are closed by pCMBS. Assuming laminar flow (within the pore), from Pos/Pd of 13 to 18 an unreasonably large pore radius of 12 to 15 A is calculated which would not hinder cell entry of known extracellular markers. Alternatively, for a single-file pore, 11 to 20 would be the number of water molecules which would be in tandem inside the pore.(ABSTRACT TRUNCATED AT 250 WORDS)
Carpi-Medina
P
P
Venezuelan Institute of Scientific Investigation, IVIC, Caracas.
León
V
V
Espidel
J
J
Whittembury
G
G
D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D001834:Body Water / Q000378:metabolism*; D002463:Cell Membrane Permeability; D004058:Diffusion; D066298:In Vitro Techniques; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000502:physiology*; D009682:Magnetic Resonance Spectroscopy; D008345:Manganese / Q000502:physiology; D008565:Membrane Proteins / Q000502:physiology; D009995:Osmosis; D011817:Rabbits; D013997:Time Factors
D008565:Membrane Proteins; D008345:Manganese; D002731:4-Chloromercuribenzenesulfonate