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PubMed1974

Osmosis in cortical collecting tubules. ADH-independent osmotic flow rectification.

The present experiments were designed to evaluate the effects of varying the osmolality of luminal solutions on the antidiuretic hormone (ADH)-independent water and solute permeability properties of isolated rabbit cortical collecting tubules. In the absence of ADH, the osmotic water permeability coefficient (cm s(-1)) P(f) (l-->b), computed from volume flows from hypotonic lumen to isotonic bath, was 20 +/- 4 x 10(-4) (SEM); the value of P(f) (b-->l) in the absence of ADH, computed from volume flows from isotonic bath to hypertonic lumen, was 88 +/- 15 x 10(-4) cm s(-1). We also measured apparent urea permeability coefficients (cm s(-1)) from (14)C-urea fluxes from lumen to bath (P(DDurea) (l-->b)) and from bath to lumen (P(DDurea) (b-->l)). For hypotonic luminal solutions and isotonic bathing solutions, P(DDurea) (l-->b) was 0.045 +/- 0.004 x 10(-4) and was unaffected by ADH. The ADH-independent values of P(DDurea) (l-->b) and P(urea) (b-->l) were, respectively, 0.216 +/- 0.022 x 10(-4) cm s(-1) and 0.033 +/- 0.002 x 10(-4) cm s(-1) for isotonic bathing solutions and luminal solutions made hypertonic with urea, i.e., there was an absolute increase in urea permeability and asymmetry of urea fluxes. Significantly, P(DDurea) (l-->b) did not rise when luminal hypertonicity was produced by sucrose; and, bathing fluid hypertonicity did not alter tubular permeability to water or to urea. We interpret these data to indicate that luminal hypertonicity increased the leakiness of tight junctions to water and urea but not sucrose. Since the value of P(f) (b-->l) in the absence of ADH, when tight junctions were open to urea, was approximately half of the value of P(f) (l-->b) in the presence of ADH, when tight junctions were closed to urea, we conclude that tight junctions are negligible paracellular shunts for lumen to bath osmosis with ADH. These findings, together with those in the preceding paper, are discussed in terms of a solubility-diffusion model for water permeation in which ADH increases water solubility in luminal plasma membranes.

Animals↗
PubMed2005

Mechanism of osmotic flow in a periodic fiber array.

The classic analysis by Anderson and Malone (Biophys J 14: 957-982, 1974) of the osmotic flow across membranes with long circular cylindrical pores is extended to a fiber matrix layer wherein the confining boundaries are the fibers themselves. The equivalent of the well-known result for the reflection coefficient sigma0 = (1 - phi)2, where phi is the partition coefficient, is derived for a periodic fiber array of hexagonally ordered core proteins. The boundary value problem for the potential energy function describing the solute distribution surrounding each fiber is solved by defining an equivalent fluid annulus in which the pressures and osmotic forces are determined. This model is of special interest in the osmotic flow of water across a capillary wall, where recent experimental studies suggest that the endothelial glycocalyx is a quasiperiodic fiber array that serves as the primary molecular sieve for plasma proteins. Results for the reflection coefficient are presented in terms of two dimensionless numbers, alpha = a/R and beta = b/R, where a and b are the solute and fiber radii, respectively, and R is the outer radius of the fluid annulus. In general, the results differ substantially from the classic expression for a circular pore because of the large difference in the shape of the boundary along which the osmotic force is generated. However, as in circular pore theory, one finds that the reflection coefficients for osmosis and filtration are the same.

Animals↗
PubMed1964

WATER TRANSFER AND CELL STRUCTURE IN ISOLATED CRAYFISH MUSCLE FIBERS.

Changes in volume of crayfish single muscle fibers in response to changes in ionic or electrical conditions have been studied in conjunction with electrophysiological measurements and electron microscopic examinations. The occurrence of at least three mechanisms of water movements is revealed, two being processes which are superimposed on the normal osmotic water movement that results from a change in the concentration of solute in the medium. Differences between the time courses of the changes in volume and potential on changing K(i)/K(o) indicate that water may be distributed unequally for a time within compartments of the fiber. Electron micrographs reveal a selective accumulation of water at the periphery of the fiber under certain conditions. A correlation of H(2)O transfer with a change in membrane potential is apparent in crayfish muscle fibers and is probably due to electroosmotic effects. Electrokinetic water movements are produced whenever the membrane potential is changed to a considerable degree by changing the level of K and/or Cl in the medium, or by applying currents with an intracellular microelectrode. Depolarizations cause shrinkage. Hyperpolarizations or repolarizations cause swelling. The volume changes are independent of the occurrence or absence of swelling in the anion-permselective transverse tubular system. They indicate that the fiber membrane along the surface is heterogeneous, not only with respect to the signs of its fixed charge sites, but also with respect to the sizes and relative permselectivities of these charged channels.

Animals↗
PubMed2005

Human red blood cells' physiological water exchange with the plasma.

In the present paper, fundamental issues related to the mechanisms of human red blood cells' physiological water exchange with the plasma (for the stationary conditions) have been discussed. It has been demonstrated, on the basis of mechanistic transport equations for membrane transport that red blood cells are capable of exchanging considerable amounts of water with the plasma. Water absorption is osmosis-driven, and its removal occurs according to the hydromechanics principle, i.e. is driven by the turgor pressure of red blood cells. This newly-acquired knowledge of these issues may appear highly useful for clinical diagnosis of blood diseases and blood circulation failures.

Animals↗
PubMed1990

Nature of the water channels in the internodal cells of Nitellopsis.

The hydraulic resistance was measured on internodal cells of Nitellopsis obtusa using the method of transcellular osmosis. The hydraulic resistance was approximately 2.65 pm-1 sec Pa, which corresponds to an osmotic permeability of 101.75 microns sec-1 (at 20 degrees C). p-Chloromercuriphenyl sulfonic acid (pCMPS) (0.1-1 mM, 60 min) reversibly increases the hydraulic resistance in a concentration-dependent manner. pCMPS does not have any effect on the cellular osmotic pressure. pCMPS increases the activation energy of water movement from 16.84 to 32.64 kJ mol-1, indicating that it inhibits water movement by modifying a low resistance pathway. pCMPS specifically increases the hydraulic resistance to exosmosis, but does not influence endosmosis. By contrast, nonyltriethylammonium (C9), a blocking agent of K+ channels, increases the hydraulic resistance to endosmosis, but does not affect that to exosmosis. These data support the hypothesis that water moves through membrane proteins in characean internodal cells and further that the polarity of water movement may be a consequence of the differential gating of membrane proteins on the endo- and exoosmotic ends.

4-Chloromercuribenzenesulfonate↗
PubMed1980

Salt-water coupling in leaky epithelia.

The theory of quasi-isotonic transport by cellular osmosis (the standing-gradient theory) has been challenged on the grounds that the osmotic permeabilities of the mucosal and interspace membranes are too low; if they were as high as the theory requires then the osmotic permeability of the whole epithelium would be 2-3 orders of magnitude higher than observed. This objection has basically been accepted for it is now claimed that these enormous permeabilities do exist, but are masked by unstirred-layer effects; I show that this is incorrect because unstirred-layer corrections are small and that the situation has not changed since 1975. The view that the route of fluid transport is junctional is replacing the cellular theory, and trans-junctional water flows seem to account for major fractions of the flow in various epithelia. I argue on grounds of general theory that these are unlikely to be osmotic flows because the junctional pores cannot satisfy both the osmotic and diffusive properties required of them, but the basic osmotic theory is also rather vague here. Non-osmotic theories, if junctional flow is accepted, must be either electro-kinetic or peristaltic.

Animals↗
PubMed1977

Osmotic properties of human lymphocyte.

The osmotic properties of human lymphocytes isolated from 15 ml of venous blood were examined. Measurements of the permeability of the membrane to water under an osmotic gradient were also made. The Boyle-Van't Hoff relation held very well for the human lymphocyte when the cells were shrunken in hyperosmotic media to concentrations twice isosmotic. The volume of osmotically inactive material or "b" value averaged 32% of the mean corpuscular volume. These values were independent of temperature. Ponder's R ranged between 0.8 and 0.9. The average value for Lp, the hydralic coefficient was 0.46 mu/min atm +/- 0.02 (S.E.M.) at 25 degrees C. No significant effect of age, sex, or race was noted. The effect of temperature between 10 degrees C and 37 degrees C was measured and heats of activation between 11.1 and 17.4 kcal/mole were calculated with a mean of 14.1 kcal/mole +/- 1.6 (S.E.M.). Concanavalin A at 10 microgram/1.5 X 10(6) lymphocytes produced blastogenesis of 25% or more of the lymphocytes without clumping, agglutination, or toxicity. The mean corpuscular volume increased by 21% after 72 hours due to an increase in the "b" value which increased by 80%. The volume of free water remained constant. Histograms of the distribution of cell volumes showed that volume changes were uniform throughout the population with no evidence of agglutination of clumping. The significance of these results is discussed in the context of membrane fluidity and the state of intracellular water.

Cell Membrane Permeability↗
PubMed1998

Structural specificity of serotonin effect on human erythrocyte fragility.

Serotonin, a neurotransmitter and vasoconstrictor, affects various cell properties. We have analyzed the importance of its structural components for its extensive effect on human erythrocyte fragility, using its O- and N-linked derivatives and related compounds. The results presented in this communication indicate that the amino group, free of adjacent negative charges, and the hydroxyl group are indispensable for the serotonin-induced increase in red blood cell fragility.

Cell Membrane Permeability↗
PubMed1994

Water permeability properties of the ovarian oocytes from Bufo arenarum and Xenopus laevis: a comparative study.

The water permeability properties of ovarian oocytes from Xenopus laevis and Bufo arenarum, a toad species found in the Buenos Aires region, were studied. We report that: (i) the water osmotic permeability (Pf, cm/sec x 10(-4)) was significantly higher in Bufo (6 degrees C = 12.3 +/- 2.4; 18 degrees C = 20.8 +/- 4.8) than in Xenopus oocytes (6 degrees C = 5.3 +/- 0.3; 18 degrees C = 6.2 +/- 1.6). The corresponding water diffusion permeability values (Pd, cm/sec x 10(-4)) were: Xenopus = 2.3 +/- 0.3 (6 degrees C) and 4.8 +/- 0.7 (18 degrees C); Bufo = 2.7 +/- 0.4 (6 degrees C) and 6.0 +/- 0.5 (18 degrees C). (ii) Amphotericin B increased the Pf and Pd values. The observed delta Pf/delta Pd ratio was not significantly different from the expected results (n = 3), after amphotericin B incorporation in both species. This means that the influence of unstirred layers and other potential artifactual compounds did not significantly affect our experimental results. (iii) Preincubation with gramicidin during 12 hr induced a clear increase in the oocyte volume. After that, a hypotonic shock only slightly increased the oocyte volume. Conversely, a hypertonic challenge induced a volume change significantly higher than the one observed in control conditions. (iv) Mercury ions did not affect the osmotic permeability in Xenopus oocytes but clearly inhibited, in a reversible way, the osmotic permeability in oocytes from B. arenarum. (v) Mercury ions did not reduce Pd values in either species.(ABSTRACT TRUNCATED AT 250 WORDS)

Amphotericin B↗
PubMed1988

Comparison of transcellular and transepithelial water osmotic permeabilities (Pos) in the isolated proximal straight tubule (PST) of the rabbit kidney.

Measurements of the water osmotic permeabilities of apical and basolateral membranes of PST cells and of the transepithelial permeability have been carried out using a very fast method with high temporal and spatial resolution. At 25 degrees C the values obtained are: 80.8 +/- 11.9 x 10(-4) cm3/s osmol cm2 of apical (luminal) surface area and 90.1 +/- 13.0 x 10(-4) cm3/s osmol cm2 of basement membrane area (no membrane invaginations taken in account). These values are higher than previously published values due to the use of a faster and more accurate volume measuring and recording system. The transepithelial water osmotic permeability at 25 degrees C is 77 +/- 11 in units of 10(-4) cm3/s osmol cm2 basement membrane area. The transcellular water osmotic permeability is 32 +/- 7 (same units), leaving a paracellular contribution of 45 +/- 10 (same units). In the presence of 2.5 mM parachloromercuribenzenesulfonate (pCMBS) the apical permeability is reduced with an incubation of 10-15 min to 23% of its control value and the basolateral permeability to 8% of its control value (after 25 min) but the transepithelial permeability is only reduced to about 1/2 of the control value. This leaves a transcellular permeability of 6 x 10(-4) cm3/s osmol cm2 of basement membrane area and a paracellular contribution of 33 +/- 6 (same units). These results indicate a significant contribution of the paracellular pathway to the transepithelial water osmotic permeabilities in PST.

4-Chloromercuribenzenesulfonate↗
PubMed1984

Thermodynamics of all-or-none water channel closure in red cells.

The relation of osmotic to diffusional water permeability of human red blood cells was compared after treating the cells with different concentrations of PCMBS (p-chloromercuribenzene sulfonate). After subtracting the PCMBS-insensitive permeability (presumably the water permeability of the lipid bilayer) from each, the ratio of osmotic to diffusional permeability remains invariant (approximately equal to 11) as more and more water channels are inhibited by increasing concentrations of PCMBS. This result implies that the channels close in an all-or-none way and suggests a two-state model. Analysis of the dependence of osmotic water permeability on PCMBS concentration in terms of the model reveals a 1:1 stoichiometry and a dissociation constant for the PCMBS/membrane receptor complex of about 0.019 mM at 37 degrees C. Temperature dependence studies show that the reaction is entropically driven (delta H degrees approximately equal to 25 kcal/mol, delta S degrees approximately equal to 100 cal/mol-deg) and suggest the involvement of hydrophobic interactions.

Cell Membrane Permeability↗
PubMed1984

Effects of potassium-free media on ADH action in toad urinary bladder.

We studied the effects of potassium-free media on processes related to the hydro-osmotic response of toad bladder to ADH (20 mU/ml). Exposure of bladders to potassium-free media did not affect base-line osmotic water flow, but it promptly attenuated the level of osmotic water permeability induced by ADH. Both the frequency of hormonally induced intra(luminal)membrane particle aggregates (presumed sites for transmembrane water flow) and the number of luminal membrane fusion events (associated with aggregate delivery from the cytoplasm) were also reduced. Potassium-free media had no measurable effect either on cytoplasmic microtubule integrity or on mean aggregate size. Potassium repletion reversed the inhibitory effect of potassium-free media on ADH-related osmotic water permeability. For bladders fully stimulated with ADH in the presence of potassium, subsequent bathing media depletion of potassium led to an inhibition of ADH-related water flow and to reductions in membrane fusion sites and aggregates. We confirmed that the inhibitory effect of potassium-free media on ADH-induced osmotic water permeability results from serosal bathing medium potassium depletion alone and occurs at a post-cyclic AMP site. In addition, we found that ADH-stimulated water permeability was attenuated in bathing media containing a low potassium concentration (0.5 mM). The data are consistent with the view that potassium-free media or media containing low levels of potassium inhibit ADH-enhanced osmotic water permeability in toad bladder by interfering with the process of or leading to membrane fusion required for the delivery of water-conducting structures to the luminal membrane. In addition, some of our results imply that aggregates may turn over during sustained ADH stimulation.

Animals↗