Determination of Equivalent Pore Radius for Human Red Cells by Osmotic Pressure Measurement

A new method has been developed to measure the equivalent pore radius in cellular membranes, and has been applied to human red cells. When red cells are suddenly introduced into a non-isosmolar concentration of non-lipid-soluble non-electrolyte molecules, water will enter or leave the cell. The rate of cell swelling or shrinking is determined and extrapolated to zero time to give the initial rate of volume change. By suitable adjustment of the concentration of the external solution the initial rate may be brought to zero. The transient equilibrium concentration, determined by interpolation from experimental data, gives a measure of Staverman's reflection coefficient, sigma. The zero time method has enabled us to determine sigma for nine permeant molecules. sigma is directly related to the equivalent pore radius; the experimental data lead to a value of 4.2 A for the equivalent pore radius in man, in good agreement with the previous figure of 3.5 A given by Paganelli and Solomon. The zero time method offers a number of advantages over previous methods for determination of this parameter. It requires no measurement of the rate of water entrance into the cell, and is essentially independent of the kinetics of cell swelling. It may be applied to a variety of living cells so that many additional membranes may now be characterized in terms of their equivalent pore radius.