THE PARACRYSTALLINE STATE OF THE RAT RED CELL

Abstract

When washed rat red cells are kept in 3 per cent sodium citrate at low temperatures (4-9 degrees C.), their resistance to osmotic hemolysis increases so that after several days they swell very little in hypotonic solutions (R = 0.15 to zero) and do not hemolyze even in distilled water. In this and in other respects they behave as if they were gelated or paracrystalline. The paracrystalline state is reversible, disappearing when the cells are warmed and rapidly reappearing when they are cooled, and the resistance to hypotonic hemolysis is not due to the cells reaching equilibrium with their environment by losing so much K that the concentrations become equal inside and out. The concentration of K remains about 25 times as great inside the cell as outside it in a hypotonic medium of T = 0.1, and the failure to swell and to hemolyze seems to be due to the activity of K in the interior of the paracrystalline cell approaching zero. The paracrystalline red cells are more resistant to saponin and digitonin hemolysis, and do not undergo the usual shape transformations, probably because they are too rigid. Hemolysis by saponin and similar lysins occurs without sphere formation, and after lysis is complete a granular debris is left behind. The paracrystalline cells show a diffuse birefringence with polarized light; on their being warmed, the birefringence disappears except at foci which are usually situated along the rim of the cell. The occurrence of the paracrystalline state accounts for the different amounts of swelling of red cells which have been observed in systems of the same degree of hypotonicity, and its relation to other metastable states of the red cell is discussed in connection with a tabulation of the metastable states of the mammalian red cell and their relation to one another. Changes in a membrane alone seem inadequate to account for the varied phenomena observed in connection with red cell behavior, the explanation of which appears to require a more detailed knowledge of the molecular architecture of the cell interior.