Ionic permeabilities of rat renal cortical brush-border membrane vesicles.
It is generally assumed that electrolytes equilibrate readily across renal cortical brush-border membrane vesicles (BBMV). This assumption was tested by use of two new methods in rat BBMV prepared with free-flow electrophoresis (FFE), Mg aggregation, or Ca aggregation. Intravesicular KCl and RbCl concentrations, as well as the conductance of Cl relative to K (GCl/GK) and GNa/GK were determined with the fluorescent, potential-sensitive probe 3,3'-dipropylthiadicarbocyanine iodide [diS-C3-(5)]; intravesicular KCl concentration was also approximated utilizing the response of Na-dependent [3H]glucose uptake to variations in the membrane potential. These studies demonstrated that KCl fails to attain electrochemical equilibrium in BBMV prepared by the three methods, despite prolonged incubation at 22 degrees C; a significant, inwardly directed electrolyte gradient was sustained in all cases. The intravesicular electrolyte concentration was lower in BBMV prepared with FFE than in those prepared with Mg or Ca. GCl/GK was lowest in BBMV prepared with FFE and highest in those prepared with Ca; GNa/GK was comparable in all preparations. The apparent impermeance of BBMV may impact significantly in interpreting data from studies that require knowledge of the precise concentration of intravesicular electrolytes.
D000818:Animals; D052160:Benzothiazoles; D002232:Carbocyanines; D002458:Cell Fractionation / Q000379:methods; D002463:Cell Membrane Permeability; D004553:Electric Conductivity; D005947:Glucose / Q000378:metabolism; D066298:In Vitro Techniques; D007672:Kidney Cortex / Q000502:physiology*; D008564:Membrane Potentials; D008871:Microvilli / Q000502:physiology; D011188:Potassium / Q000378:metabolism; D051381:Rats; D012413:Rubidium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism
D052160:Benzothiazoles; D002232:Carbocyanines; C012944:3,3'-dipropyl-2,2'-thiadicarbocyanine; D012964:Sodium; D005947:Glucose; D012413:Rubidium; D011188:Potassium