scholarly journals An Analysis of Unstirred Layers in Series with "Tight" and "Porous" Lipid Bilayer Membranes

1971 ◽  
Vol 57 (4) ◽  
pp. 464-478 ◽  
Author(s):  
Thomas E. Andreoli ◽  
Susan L. Troutman

The present experiments were designed to evaluate the effective thickness of the unstirred layers in series with native and porous (i.e., in the presence of amphotericin B) lipid bilayer membranes and, concomitantly, the respective contributions of membranes and unstirred layers to the observed resistances to the diffusion of water and nonelectrolytes between aqueous phases. The method depended on measuring the tracer permeability coefficients for the diffusion of water and nonelectrolytes (PDDi, cm sec-1) when the aqueous phase viscosity (η) was increased with solutes having a unity reflection coefficient, such as sucrose or dextran. The effective thickness of the unstirred layers (αt, cm) and the true, or membrane, permeability coefficients for diffusion of water and nonelectrolytes (Pmmi, cm sec-1) were computed from, respectively, the slope and intercept of the linear regression of 1/PDDi on η. In both the native and porous membranes, αt was approximately 110 x 10-4 cm. The ratio of Pf, the osmotic water permeability coefficient (cm sec-1) to PmmH2O was 1.22 in the native membranes and 3.75 in the porous membranes. For the latter, the effective pore radius, computed from Poiseuille's law, was approximately 5.6 A. A comparison of Pmmi and PDDi, indicated that the porous membranes accounted for 16, 25, and 66% of the total resistance to the diffusion of, respectively, H2O, urea, and glycerol, while the remainder was referable to the unstirred layers.

1971 ◽  
Vol 57 (4) ◽  
pp. 479-493 ◽  
Author(s):  
Thomas E. Andreoli ◽  
James A. Schafer ◽  
Susan L. Troutman

The present experiments were designed to evaluate coupling of water and nonelectrolyte flows in porous lipid bilayer membranes (i.e., in the presence of amphotericin B) in series with unstirred layers. Alterations in solute flux during osmosis, with respect to the flux in the absence of net water flow, could be related to two factors: first, changes in the diffusional component of solute flux referable to variations in solute concentrations at the membrane interfaces produced by osmotic flow through the unstirred layers; and second, coupling of solute and solvent flows within the membrane phase. Osmotic water flow in the same direction as solute flow increased substantially the net fluxes of glycerol and erythritol through the membranes, while osmotic flow in the opposite direction to glycerol flow reduced the net flux of that solute. The observed effects of osmotic water flow on the fluxes of these solutes were in reasonable agreement with predictions based on a model for coupling of solute and solvent flows within the membrane phase, and considerably in excess of the prediction for a diffusion process alone.


1974 ◽  
Vol 64 (2) ◽  
pp. 201-228 ◽  
Author(s):  
James A. Schafer ◽  
Clifford S. Patlak ◽  
Thomas E. Andreoli

This paper reports a theoretical analysis of osmotic transients and an experimental evaluation both of rapid time resolution of lumen to bath osmosis and of bidirectional steady-state osmosis in isolated rabbit cortical collecting tubules exposed to antidiuretic hormone (ADH). For the case of a membrane in series with unstirred layers, there may be considerable differences between initial and steady-state osmotic flows (i.e., the osmotic transient phenomenon), because the solute concentrations at the interfaces between membrane and unstirred layers may vary with time. A numerical solution of the equation of continuity provided a means for computing these time-dependent values, and, accordingly, the variation of osmotic flow with time for a given set of parameters including: Pf (cm s–1), the osmotic water permeability coefficient, the bulk phase solute concentrations, the unstirred layer thickness on either side of the membrane, and the fractional areas available for volume flow in the unstirred layers. The analyses provide a quantitative frame of reference for evaluating osmotic transients observed in epithelia in series with asymmetrical unstirred layers and indicate that, for such epithelia, Pf determinations from steady-state osmotic flows may result in gross underestimates of osmotic water permeability. In earlier studies, we suggested that the discrepancy between the ADH-dependent values of Pf and PDDw (cm s–1, diffusional water permeability coefficient) was the consequence of cellular constraints to diffusion. In the present experiments, no transients were detectable 20–30 s after initiating ADH-dependent lumen to bath osmosis; and steady-state ADH-dependent osmotic flows from bath to lumen and lumen to bath were linear and symmetrical. An evaluation of these data in terms of the analytical model indicates: First, cellular constraints to diffusion in cortical collecting tubules could be rationalized in terms of a 25-fold reduction in the area of the cell layer available for water transport, possibly due in part to transcellular shunting of osmotic flow; and second, such cellular constraints resulted in relatively small, approximately 15%, underestimates of Pf.


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