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The role of water in protoplasmic permeability and in antagonism.

The behavior of the cell depends to a large extent on the permeability of the outer non-aqueous surface layer of the protoplasm. This layer is immiscible with water but may be quite permeable to it. It seems possible that a reversible increase or decrease in permeability may be due to a corresponding increase or decrease in the water content of the non-aqueous surface layer. Irreversible increase in permeability need not be due primarily to increase in the water content of the surface layer but may be caused chiefly by changes in the protoplasm on which the surface layer rests. It may include desiccation, precipitation, and other alterations. An artificial cell is described in which the outer protoplasmic surface layer is represented by a layer of guaiacol on one side of which is a solution of KOH + KCl representing the external medium and on the other side is a solution of CO(2) representing the protoplasm. The K(+) unites with guaiacol and diffuses across to the artificial protoplasm where its concentration becomes higher than in the external solution. The guaiacol molecule thus acts as a carrier molecule which transports K(+) from the external medium across the protoplasmic surface. The outer part of the protoplasm may contain relatively few potassium ions so that the outwardly directed potential at the outer protoplasmic surface may be small but the inner part of the protoplasm may contain more potassium ions. This may happen when potassium enters in combination with carrier molecules which do not completely dissociate until they reach the vacuole. Injury and recovery from injury may be studied by measuring the movements of water into and out of the cell. Metabolism by producing CO(2) and other acids may lower the pH and cause local shrinkage of the protoplasm which may lead to protoplasmic motion. Antagonism between Na(+) and Ca(++) appears to be due to the fact that in solutions of NaCl the surface layer takes up an excessive amount of water and this may be prevented by the addition of suitable amounts of CaCl(2). In Nitella the outer non-aqueous surface layer may be rendered irreversibly permeable by sharply bending the cell without permanent damage to the inner non-aqueous surface layer surrounding the vacuole. The formation of contractile vacuoles may be imitated in non-living systems. An extract of the sperm of the marine worm Nereis which contains a highly surface-active substance can cause the egg to divide. It seems possible that this substance may affect the surface layer of the egg and cause it to take up water. A surface-active substance has been found in all the seminal fluids examined including those of trout, rooster, bull, and man. Duponol which is highly surface-active causes the protoplasm of Spirogyra to take up water and finally dissolve but it can be restored to the gel state by treatment with Lugol solution (KI + I). The transition from gel to sol and back again can be repeated many times in succession. The behavior of water in the surface layer of the protoplasm presents important problems which deserve careful examination.

Animals↗

Water permeability of thin lipid membranes.

The osmotic permeability coefficient, P(f), and the tagged water permeability coefficient, P(d), were determined for thin (<100 A) lipid membranes formed from ox brain lipids plus DL-alpha-tocopherol; their value of approximately 1 x 10(-3) cm/sec is within the range reported for plasma membranes. It was established that P(f) = P(d). Other reports that P(f) > P(d) can be attributed to the presence of unstirred layers in the experimental determination of P(d). Thus, there is no evidence for the existence of aqueous pores in these thin phospholipid membranes. The adsorption onto the membrane of a protein that lowers its electrical resistance by a factor of 10(3) was found not to affect its water permeability; however, glucose and sucrose were found to interact with the membrane to modify P(f). Possible mechanisms of water transport across these films are discussed, together with the implications of data obtained on these structures for plasma membranes.

Adsorption↗

Permeability of Serratia marcescens to some inorganic salts.

The physical interactions between Serratia marcescens and solutions of NaCl, CaCl(2), CaI(2), NaI, and Na(2)HPO(4) plus NaH(2)PO(4) were examined. Dilute (0.017 n) salt solutions did not cause cells to lose water, as evidenced by the unchanged weight of centrifugally packed cells. The cells preferentially adsorbed the cations and repelled the anions of most salts in these solutions. Concentrated (1.71 n) salt solutions markedly reduced the weight and water content of centrifugally packed cells, although these cells took up considerable amounts of salts. More than 90% of the water in the packed-cell pellets was available for the solution of NaCl at 4.2 to 4.4% concentration. The observation that salts apparently penetrated the cells freely and yet caused extensive dehydration was not readily compatible with conventional concepts of solute-induced plasmolysis. Alternative hypotheses to explain the data included the following. First, the cells lost weight and water to concentrated salt solutions through a nonosmotic competitive dehydration, causing a shrinkage of the protoplasmic gel. The shrinkage of the cell wall was limited because of the rigidity of its mucopeptide layer; therefore, a space appeared between the cell wall and the cell membrane. Second, cells may have equilibrated their water activity with that of their environment by two mechanisms: (i) the loss of water by plasmolysis or competitive dehydration, and (ii) alterations in cell permeability that admitted previously excluded solutes to the cell interior. Possibly, the correct explanation of the observations reported here involves elements of all three hypotheses, plasmolysis, competitive dehydration, and permeability alterations.

Calcium Chloride↗

Osmotic shrinkage of giant egg-lecithin vesicles.

Osmotic shrinkage of giant egg-lecithin vesicles was observed by phase-contrast microscopy. The vesicles remained or became spherical when shrinking. Small and thick-walled vesicles formed visible fingers attached to the sphere. The water permeability of the single bilayer was found to be 41 micrometers/s. A variety of observations indicate that osmosis induces a parallel lipid flow between the monolayers of the bilayer, leading to a strong positive spontaneous curvature. They also suggest the formation of mostly submicroscopic daughter vesicles. The estimated coupling constant, 2 . 10(-6) mol/mol, is large enough to be biologically significant.

Lipid Bilayers↗

Mechanistic formalism for membrane transport generated by osmotic and mechanical pressure.

Since the physical interpretation of practical Kedem-Katchalsky (KK) equations is not clear, we consider an alternative, mechanistic approach to membrane transport generated by osmotic and hydraulic pressure. We study a porous membrane with randomly distributed pore sizes (radii). We postulate that reflection coefficient (sigma p) of a single pore may equal 1 or 0. From this postulate we derive new (mechanistic) transport equations. Their advantage is in clear physical interpretation and since we show they are equivalent to the KK equations, the interpretation of the latter became clearer as well. Henceforth the equations allow clearer and more detailed interpretation of results concerning membrane mass transport. This is especially important from the point of view of biophysical studies on permeation processes across biological membranes, cell membranes including.

Biological Transport↗