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Some physical parameters controlling cell size during the evolution of the procaryons.

Possible factors controlling cell size during the evolution of unicellular organisms have been examined. It has been shown that considerations of osmotic and membrane pressure eqilibria will predict minimal cell sizes which are in good agreement with those found in present day microorganisms. It has also been shown that the possibility of random proton 'noise' would not be a limiting factor for even the smallest organisms or structures. Maximum cell size would be governed by the requirements of diffusion and transport within the cell.

Bacteria↗

Osmotic water permeability of plasma and vacuolar membranes in protoplasts II: theoretical basis.

Water permeability of the plasma membrane (PM) and the vacuolar membrane (VM) is important for intracellular and transcellular water movement in plants, because mature plant cells have large central vacuoles. We have developed a new method for measuring the osmotic water permeability of the PM and VM (P ( f1) and P ( f2), respectively) in individual plant cells. Here, the theoretical basis and procedure of the method are discussed. Protoplasts isolated from higher plant tissues are used to measure P ( f1) and P ( f2). Because of the semi-permeability (selective permeability) of cellular membranes, protoplasts swell or shrink under hypotonic or hypertonic conditions. A theoretical three-compartment model is presented for simulating time-dependent volume changes in the vacuolar and cytoplasmic spaces in a protoplast during osmotic excursions. The model describes the theoretical relationships between P ( f1), P ( f2) and the bulk osmotic water permeability of protoplasts (P ( f(bulk))). The procedure for measuring the osmotic water permeability is: (1) P ( f(bulk)) is calculated from the time when half of the total change in protoplast volume is completed, by assuming that the protoplast has a single barrier to water movement across it (two-compartment model); (2) P ( f2) of vacuoles isolated from protoplasts is obtained in the same manner; and (3) P ( f1) is determined from P ( f(bulk)) and P ( f2) according to the three-compartment model. The theoretical relationship between P ( fl ) (m s(-1)) and L ( Pl ) (hydraulic conductivity, l=1, 2) (m s(-1) Pa(-1)) is also discussed.

Cell Membrane Permeability↗

The failure of hydrodynamic analysis to define pore size in cell membranes.

The equivalent pore theory predicts that the size of water transporting pores can be calculated from the ratio of osmotic (Pf, cm . s-1) to diffusive (Pd, cm . s-1) water permeability. Determinations of Pf and Pd in human red cells within the last thirty years have increased the ratio of Pf to Pd. According to the equivalent pore theory the pore diameter has increased from 9 A to 25 A. A pore diameter of 25 A is not compatible with the permeability characteristics of the red cell membrane. We conclude that the equivalent pore theory fails to determine pore size in red blood cells. We suggest that water transporting pores in human red cells transport water molecules in a single file fashion.

4-Chloromercuribenzenesulfonate↗

The osmotic permeability of isolated calf pulmonary artery endothelial cells.

Osmotic permeability coefficients, PF, for water in isolated calf pulmonary artery endothelial cells determined over the temperature range 41 to 20 degrees C are 311.10(-5) cm.s-1 at 37 degrees C and 159.10(-5) cm.s-1 at 20 degrees C. The value at 37 degrees C is close to that reported earlier for the diffusional permeability coefficient, PD. The PF/PD ratio is 1.0 at 37 degrees C. The PF values are within the range of values extrapolated for filtration permeability in pulmonary endothelium. The temperature dependence expressed as the activation energy is 7.2 kcal.mol-1. The product of hydraulic conductivity, Lp (or PF) and of viscosity changes in water is not constant from 37 to 20 degrees C. These results can be interpreted to indicate a similar pathway for water whether under diffusional or osmotic gradients.

Animals↗

Differential blood-brain barrier permeabilities to [14C]sucrose and [3H]inulin after osmotic opening in the rat.

The blood-brain barrier (B-BB) in 3-month-old rats was opened unilaterally by infusing 1.8 m L(+)arabinose in water into the internal carotid artery through a catheter in the external carotid. Two poorly penetrating uncharged test radiotracers of differing molecular weight and size, [14C]sucrose (340 daltons, radius 5 A) and [3H]inulin (5500 daltons, radius 15 A), were simultaneously injected i.v. in untreated rats, or rats at 1, 30, or 50 min after infusion of hypertonic arabinose solution. Evans-blue solution was injected 5 min prior to osmotic treatment as a visual indicator of barrier integrity. In regions of uninfused control brains, the [14C]sucrose permeability-surface area (PA) product approximated 10(-5) s-1, whereas PA was not measurable for [3H]inulin. In arabinose-infused animals, PA products on the ipsilateral hemisphere for both [14C]sucrose and [3H]inulin were markedly elevated 6 min after infusion, but decreased by 35 and 55 min. In nearly all regions, statistically significant differences were not found between 6-min [14C]sucrose- and [3H]inulin-PA values (P greater than 0.05). However, at 35 and 55 min in most regions, the PA for [3H]inulin was significantly lower (P less than 0.05) than PA for [14C]sucrose. The results indicated that the B-BB closed more rapidly to larger than to smaller molecules after osmotic treatment and were consistent with a pore model for osmotic B-BB opening.

Animals↗

Analytical solution for the extremums of cell water volume and cell volume using a two-parameter model.

Based on a two-parameter model [Cryobiology 37 (1998) 271-289], the analytical solution for the extremums of cell water volume and cell volume for a two-solute system are obtained. Compared with the numerical solution, the analytical solution offers an accurate but simple choice. The approximate solution [Cryobiology 40 (2000) 64-83] for the extremum of cell water volume is also discussed, the reason for the deviation is presented.

Cell Membrane Permeability↗

Further observations on asymmetrical solute movement across membranes.

The permeability of frog skin under the influence of urea hyperosmolarity has been studied. Flux ratio asymmetry has been demonstrated again for tracer mannitol. The inhibitors DNP, CN(-), and ouabain have been used to eliminate active sodium transport and it was found that urea hyperosmolarity produces asymmetrical mannitol fluxes on frog skins having no short-circuit current. These findings suggest that flux ratio asymmetry is due to solute interaction and is unrelated to sodium transport. Studies with a synthetic membrane show clearly that bulk flow of fluid can produce a "solvent drag" effect and change flux ratios. When bulk flow is blocked and solute gradients allowed their full expression, then solute interaction "solute drag" is easily demonstrable in a synthetic system.

Animals↗

Further quantification of the role of internal unstirred layers during the measurement of transport coefficients in giant internodes of Chara by a new stop-flow technique.

A new stop-flow technique was employed to quantify the impact of internal unstirred layers on the measurement of the solute permeability coefficient (P(s)) across the plasma membrane of internodes of the giant-celled alga Chara corallina using a cell pressure probe. During permeation experiments with rapidly permeating solutes (acetone, 2-propanol, and dimethylformamide), the solute concentration inside the cell was estimated and the external medium was adjusted to stop solute transport across the membrane, after which responses in turgor were measured. This allowed estimation of the solute concentration right at the membrane. Stop-flow experiments were also simulated with a computer. Both the stop-flow experiments and simulations provided quantitative data about internal concentration gradients and the contribution of unstirred layers to overall measured values of P(meas)(s) for the three solutes. The stop-flow experimental results agreed with stop-flow simulations assuming that solutes diffused into a completely stagnant cell interior. The effects of internal unstirred layers on the underestimation of membrane P(s) declined with decreasing P(s). They were no bigger than 37% in the presence of the most rapidly permeating solute, acetone (P(meas)(s) =4.2 x 10(-6) m s(-1)), and 14% for the less rapidly permeating dimethylformamide (P(meas)(s) =1.6x10(-6) m s(-1)). It is concluded that, even in the case of rapidly permeating solutes such as isotopic water and, even when making pessimistic assumptions about the internal mixing of solutes, an upper limit for the underestimation of P(s) due to internal unstirred layers was 37%. The data are discussed in terms of recent theoretical estimates of the effect of internal unstirred layers and in terms of some recent criticism of cell pressure probe measurements of water and solute transport coefficients. The current stop-flow data are in line with earlier estimations of the role of unstirred layers in the literature on cell water relations.

Biological Transport↗

Water and solute permeability of rat lung caveolae: high permeabilities explained by acyl chain unsaturation.

Caveolae are invaginated membrane structures with high levels of cholesterol, sphingomyelin, and caveolin protein that are predicted to exist as liquid-ordered domains with low water permeability. We isolated a caveolae-enriched membrane fraction without detergents from rat lung and characterized its permeability properties to nonelectrolytes and protons. Membrane permeability to water was 2.85 +/- 0.41 x 10(-3) cm/s, a value 5-10 times higher than expected based on comparisons with other cholesterol and sphingolipid-enriched membranes. Permeabilities to urea, ammonia, and protons were measured and found to be moderately high for urea and ammonia at 8.85 +/- 2.40 x 10(-7)and 6.84 +/- 1.03 x 10(-2) respectively and high for protons at 8.84 +/- 3.06 x 10(-2) cm/s. To examine whether caveolin or other integral membrane proteins were responsible for high permeabilities, liposomes designed to mimic the lipids of the inner and outer leaflets of the caveolar membrane were made. Osmotic water permeability to both liposome compositions were determined and a combined inner/outer leaflet water permeability was calculated and found to be close to that of native caveolae at 1.58 +/- 1.1 x 10(-3) cm/s. In caveolae, activation energy for water flux was high (19.4 kcal/mol) and water permeability was not inhibited by HgCl2; however, aquaporin 1 was detectable by immunoblotting. Immunostaining of rat lung with AQP1 and caveolin antisera revealed very low levels of colocalization. We conclude that aquaporin water channels do not contribute significantly to the observed water flux and that caveolae have relatively high water and solute permeabilities due to the high degree of unsaturation in their fatty acyl chains.

Ammonia↗

Human platelet osmotic water and nonelectrolyte transport.

The osmotic water (Pf) and nonelectrolyte permeability (Ps) properties of human platelets were characterized using the stopped-flow light-scattering technique. At 37 degrees C, Pf = 0.007 +/- 0.001 cm/s, the urea reflection coefficient (sigma urea) = 0.95 +/- 0.04, and Ps for a series of permeant nonelectrolytes was (in cm X s-1 X 10(-6)) 2.1 (urea), 3.5 (glycerol), 3.8 (thiourea), 17 (ethylene glycol), 18 (acetamide), 23 (formamide), and 24 (butyramide). Pf did not depend on the size of the osmotic gradient or on the direction of volume flow. Mercurial sulfhydryl reagents did not inhibit osmotic water transport, and phloretin and phenylurea did not inhibit urea transport. There was a discontinuity in the temperature dependence for both Pf and urea permeability (P urea) at 36 degrees C; enthalpy (delta H) = 25 (greater than 36 degrees C) and 4.4 kcal/mol (less than 36 degrees C) for Pf, and delta H = 26 (greater than 36 degrees C) and 7 kcal/mol (less than 36 degrees C) for P urea. In contrast to the facilitated water and urea transport systems in the red blood cell, these results suggest that the mechanism for water and urea transport in the platelet is primarily by diffusion through membrane phospholipid. A computer-simulated model of platelet circulation through the renal medulla, based on the measured values for Pf, P urea, and sigma urea, indicated that platelets undergo an approximately 40% decrease in volume in the inner medulla and an approximately 20% overshoot in volume as they return to the external isosmotic environment.

Amides↗

Human platelet diffusional water permeability measured by nuclear magnetic resonance.

To examine whether water transport in human platelets is mediated by an aqueous pore or channel, the ratio of osmotic to diffusional water permeability coefficients (Pf/Pd) was measured. Pd was measured from protein spin-lattice relaxation times in a dense platelet suspension (approximately 20% intracellular exchangeable water) using 20 mM solution Mn as a paramagnetic quencher. The decay of magnetization was biexponential with time constants of 1.3 and 6.9 ms (10 MHz, 37 degrees C), corresponding to a Pd of (2.9 +/- 0.2) X 10(-3) cm/s (SE; n = 8). Pd did not depend on concentrations of Mn (6-20 mM) or of platelets (2-4 X 10(10) platelets/ml), but increased to 4.5 X 10(-3) cm/s with addition of gramicidin (6 micrograms/10(10) platelets). 54Mn uptake studies showed less than 1% of equilibrium uptake of Mn into platelets in 30 min at 37 degrees C. The activation energies (Ea) for Pd were 4.5 kcal/mol (less than 28 degrees C) and 16.3 kcal/mol (greater than 28 degrees C). Pf was measured by a stopped-flow light scattering technique as reported previously [M. M. Meyer and A. S. Verkman, Human platelet osmotic water and nonelectrolyte transport, Am. J. Physiol. 251 (Cell Physiol. 20): C549-C557, 1986], in which the time course of platelet volume was measured in response to a 100 mM inwardly directed sucrose gradient. At 37 degrees C, Pf was (2.7 +/- 0.2) X 10(-3) cm/s and independent of [Mn]. The measured platelet Pf/Pd of 0.93 +/- 0.1 suggests that unlike water transport in erythrocytes, platelet water transport is not associated with an aqueous channel.

Blood Platelets↗