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PubMed1993

Transport of fluid and ions through a porous-permeable charged-hydrated tissue, and streaming potential data on normal bovine articular cartilage.

Using the triphasic mechano-electrochemical theory [Lai et al., J. biomech. Engng 113, 245-258 (1991)], we analyzed the transport of water and ions through a finite-thickness layer of charged, hydrated soft tissue (e.g. articular cartilage) in a one-dimensional steady permeation experiment. For this problem, we obtained numerically the concentrations of the ions, the strain field and the fluid and ion velocities inside when the specimen is subject to an applied mechanical pressure and/or osmotic pressure across the layer. The relationships giving the dependence of streaming potential and permeability on the negative fixed charge density (FCD) of the tissue were derived analytically for the linear case, and calculated for the nonlinear case. Among the results obtained were: (1) at a fluid pressure difference of 0.1 MPa across the specimen layer, there is a 10% flow-induced compaction at the downstream boundary; (2) the flow-induced compaction causes the FCD to increase and the neutral salt concentration to decrease in the downstream direction; (3) while both ions move downstream, relative to the solvent (water), the anions (Cl-) move with the flow whereas cations (Na+) move against the flow. The difference in ion velocities depends on the FCD, and this difference attained a maximum at a physiological FCD of around 0.2 meq ml-1; (4) the apparent permeability decreases nonlinearly with FCD, and the apparent stiffness of the tissue increases with FCD; and (5) the streaming potential is not a monotonic function of the FCD but rather it has a maximum value within the physiological range of FCD for articular cartilage. Finally, experimental data on streaming potential were obtained from bovine femoral cartilage. These data support the triphasic theoretical prediction of non-monotonicity of streaming potential as a function of the FCD.

Animals↗
PubMed1996

Osmotic and pH transmembrane gradients control the lytic power of melittin.

Transmembrane osmotic gradients applied on large unilamellar 1-palmitoyl-2-oleoyl-phosphatidylcholine vesicles were used to modulate the potency of melittin to induce leakage. Melittin, an amphipathic peptide, changes the permeability of vesicles, as studied using the release of entrapped calcein, a fluorescent marker. A promotion of the ability of melittin to induce leakage was observed when a hyposomotic gradient (i.e., internal salt concentration higher than the external one) was imposed on the vesicles. It is proposed that structural perturbations caused by the osmotic pressure loosen the compactness of the outer leaflet, which facilitates the melittin-induced change in membrane permeability. Additionally, we have shown that this phenomenon is not due to enhanced binding of melittin to the vesicles using intrinsic fluorescence of the melittin tryptophan. Furthermore, we investigated the possibility of using a transmembrane pH gradient to control the lytic activity of melittin. The potency of melittin in inducing release is known to be inhibited by increased negative surface charge density. A transmembrane pH gradient causing an asymmetric distribution of unprotonated palmitic acid in the bilayer is shown to be an efficient way to modulate the lytic activity of melittin, without changing the overall lipid composition of the membrane. We demonstrate that the protective effect of negatively charged lipids is preserved for asymmetric membranes.

Biophysical Phenomena↗
PubMed1975

Nonelectrolyte permeability of the paracellular pathway in Necturus proximal tubule.

Micropuncture experiments were performed on Necturus proximal tubule using stationary microperfusion and microrecollection techniques. The transepithelial movement of the extracellular marker, sucrose, was used to investigate the passive permeability of the paracellular shunt pathway under steady-state conditions, during spontaneous reabsorption and water flow induced by an external osmotic gradient. Measurements were made of the sucrose permeability (P-s) efflux, net flux, and of net volume flow. True P-s determined in the absence of net volume flow and transepithelial gradient was 0.96 10-6 cm s-1. Both ouabain and isotonic volume expansion decreased shunt P-s. During reabsorption, solute-coupled water flow increased apparent P-s and net sucrose flux equalled efflux. Osmotic water flow from lumen to plasma decreased apparent P-s, with net sucrose flux equal to efflux; whereas osmotic flow from plasma to lumen increased apparent P-s but no net flux was observed. It is concluded that changes in P-s can be interpreted as relative alterations of the tight junction and the lateral spaces and that a portion of the volume flow from lumen to plasma proceeds via the tight junction.

Animals↗
PubMed2005

Water relations of the pine exine.

BACKGROUND AND AIMS: Water adhesion forces, water absorption capacity and permeability of the pine exine were investigated to consider a possible function of sporopollenin coatings in the control of water transport. METHODS: The experiments were carried out with sporopollenin capsules obtained from pine pollen consisting of an empty central capsule and two sacci. Changes in the concentration of excluded dextran molecules in the medium were analysed to quantify water absorption by purified exine fragments and the osmotic volume flow out of the intact central capsule. KEY RESULTS: The contact angle of sporopollenin to water is higher than the one to ethanol and lower than the one to n-heptane. The water-filled pore space in pine sporopollenin amounts to only 20.6 % of the matrix volume. A monosaccharide was excluded from 15 % and a trisaccharide from about 38 % of this space. Shrinkage of the central capsule induced by permeable osmotica was transient, whereas that induced by sodium polyacrylate (2100 g mol(-1)) was stable. Values obtained for the hydraulic conductance L(P) of the exine (0.39-0.48 microm s(-1) MPa(-1)) are comparable in size to those of biomembranes. Sodium sulfate solutions induced a significant osmotic flow through the exine (reflection coefficient at least 0.6). The exine around the central capsule can be ruptured by equilibration of its lumen with a concentrated electrolyte solution and subsequent transfer to water. The denatured protoplast along with the intact intine was ejected when pollen grains were subjected to this osmotic shock treatment. CONCLUSIONS: The pine exine is easily wetted with water and does not represent a significant barrier to water exchange either liquid or gaseous. Through osmotic burst, it can be separated from the intine. The effect of salts and small solute molecules on water fluxes may be functionally significant for rehydration upon pollination.

Biological Transport↗
PubMed1974

Osmosis in cortical collecting tubules. A theoretical and experimental analysis of the osmotic transient phenomenon.

This paper reports a theoretical analysis of osmotic transients and an experimental evaluation both of rapid time resolution of lumen to bath osmosis and of bidirectional steady-state osmosis in isolated rabbit cortical collecting tubules exposed to antidiuretic hormone (ADH). For the case of a membrane in series with unstirred layers, there may be considerable differences between initial and steady-state osmotic flows (i.e., the osmotic transient phenomenon), because the solute concentrations at the interfaces between membrane and unstirred layers may vary with time. A numerical solution of the equation of continuity provided a means for computing these time-dependent values, and, accordingly, the variation of osmotic flow with time for a given set of parameters including: P(f) (cm s(-1)), the osmotic water permeability coefficient, the bulk phase solute concentrations, the unstirred layer thickness on either side of the membrane, and the fractional areas available for volume flow in the unstirred layers. The analyses provide a quantitative frame of reference for evaluating osmotic transients observed in epithelia in series with asymmetrical unstirred layers and indicate that, for such epithelia, P(f) determinations from steady-state osmotic flows may result in gross underestimates of osmotic water permeability. In earlier studies, we suggested that the discrepancy between the ADH-dependent values of P(f) and P(DDw) (cm s(-1), diffusional water permeability coefficient) was the consequence of cellular constraints to diffusion. In the present experiments, no transients were detectable 20-30 s after initiating ADH-dependent lumen to bath osmosis; and steady-state ADH-dependent osmotic flows from bath to lumen and lumen to bath were linear and symmetrical. An evaluation of these data in terms of the analytical model indicates: First, cellular constraints to diffusion in cortical collecting tubules could be rationalized in terms of a 25-fold reduction in the area of the cell layer available for water transport, possibly due in part to transcellular shunting of osmotic flow; and second, such cellular constraints resulted in relatively small, approximately 15%, underestimates of P(f).

Animals↗
PubMed2003

Angiotensin-(1-7) stimulates water transport in rat inner medullary collecting duct: evidence for involvement of vasopressin V2 receptors.

The peptide angiotensin-(1-7) [Ang-(1-7)] is known to enhance water transport in rat inner medullary collecting duct (IMCD). The aim of this study was to determine the mechanism of the Ang-(1-7) effect on osmotic water permeability (Pf). Pf was measured in the normal rat IMCD perfused in vitro in presence of agonists [Ang-(1-7), arginine vasopressin (AVP) and Ang-(3-8)], and antagonists of the angiotensin and the vasopressin cascade. Ang-(1-7), but not Ang-(3-8), increased Pf significantly. The effect of Ang-(1-7) on Pf was abolished by its selective antagonist, A-779, added before or after Ang-(1-7). Prostaglandin E2 and the protein kinase A inhibitor H8 also blocked the Ang-(1-7) effect. Blockade of vasopressin V1 receptors by antagonists did not change the Ang-(1-7) effect, but pre-treatment with a V2 antagonist abolished the effect of Ang-(1-7) on Pf. Similarly, pre-treatment with A-779 inhibited AVP's effect on Pf. Forskolin-stimulated Pf was blocked both by A-779 and by the V2 antagonist. Finally, Ang-(1-7) increased cAMP levels in fresh IMCD cell suspensions whilst the forskolin-stimulated cAMP synthesis was decreased by A-779 and the V2 antagonist. These data provide evidence that Ang-(1-7) interacts via its receptor with the AVP V2 system through a mechanism involving adenylate-cyclase activation.

Angiotensin I↗
PubMed1958

Water transport in invertebrate peripheral nerve fibers.

Osmotic and diffusion permeabilities (P(f) and P(d)) of invertebrate nerve fibers to tritiated water were measured to determine what water flux studies could reveal about "the nerve membrane" and to directly test the possibility of active transport of water into or out of invertebrate nerve fibers. P(f)/P(d) ratios for lobster walking leg nerve fibers were found to be about 20 +/- 7 at 14 degrees C. P(d) measurements were made for squid giant axons at 25 degrees C. and found to yield a value of 4 x 10(-4) cm.(-1) sec.(-1). When combined with the data of D. K. Hill for P(f), a P(f)/P(d) ratio of 21 +/- 5 is obtained. These P(f)/P(d) ratios correspond to "effective pore radii" of about 16 +/- 4 angstrom units, according to theories developed by Koefoed-Johnsen and Ussing and independently by Pappenheimer and his colleagues. Variations of water flux ratios with temperatures were studied and apparent activation energies calculated for both diffusion experiments and osmotic filtration experiments using the Arrhenius equation, and found to be close to 3 to 5 cal. per mole of water transferred. Cyanide (5 x 10(-3) molar) and iodoacetate (1 x 10(-3) molar) poisoned lobster leg nerve fibers showed no appreciable change in diffusion or osmotic filtration water effluxes. Caution in interpreting these proposed channels as simple pores was emphasized, but the possibility that such channels exist and are related to ionic flow is not incompatible with electrophysiological data.

Biological Transport↗
PubMed1959

Isotonicity of liver and of kidney tissue in solutions of electrolytes.

Solutions of a wide variety of electrolytes, isotonic with liver or with kidney tissue, have approximately the same osmotic pressure as solutions of sodium chloride isotonic with tissues of the two organs respectively; that is, with solutions approximately twice as concentrated as the sodium chloride of mammalian blood plasma. The molar concentration of various electrolytes isotonic with liver or with kidney tissue immediately after its removal from the body is determined by the molecular weight, valency, and ion-dissociation of these electrolytes in accordance with the well known conditions of osmosis. The plasma membranes of liver and of kidney cells are imperfectly semipermeable to electrolytes, and those that enter the cell, though retarded in so doing, bring about injury which increases permeability to water. The osmotic activity of cells of mammalian liver and kidney immediately after their removal from the body resembles that of plant cells, egg cells of marine invertebrates, and mammalian red blood corpuscles and presumably represents a basic property of living cells by which osmotic pressure may be adjusted to functional need.

Animals↗
PubMed1976

Comparative aspects of salt and water transport across lung.

Calculation suggests that the outward flux of water from the surface of the lung into the expired air could be driven by a relatively small standing osmotic gradient. The structure of this alveolar epithelium does not appear to possess the characteristics of a typical secretory epithelium. These considerations argue against the occurrence of active transport in the alveoli of the adult mammalian lung.

Animals↗
PubMed2005

Transdermal iontophoretic delivery of triptorelin in vitro.

The feasibility of delivering triptorelin ([D-Trp6]LHRH) by transdermal iontophoresis was evaluated in vitro. Peptide electrotransport at different current densities and donor concentrations was measured across porcine ear skin. The concomitant delivery of an electroosmotic marker enabled calculation of the respective contributions of electromigration (EM) and electroosmosis (EO) to iontophoretic delivery. At a given concentration (3 mM), a threefold increase in current density produced a corresponding increase in the cumulative amount of peptide present in the receptor compartment. Conversely, doubling the concentration to 6 mM produced a twofold reduction in the amount of peptide delivered, partly due to a concentration-dependent inhibition of EO. EM was revealed to be the predominant transport mechanism, accounting for 80% of overall delivery. Finally, despite the inhibition of EO, the results indicate that application of an iontophoretic current of 0.8 mA over a relatively small contact area (4 cm2) would provide a delivery rate of 36 microg/h, largely sufficient for therapeutic requirements.

Acetaminophen↗
PubMed1981

Barriers to water flow in vasopressin-treated toad urinary bladder.

Unstirred layers of water complicate the measurement of water permeability across epithelia. In the toad urinary bladder, the hormone vasopressin increases the osmotic water permeability of the granular epithelial cell's luminal membrane, and also leads to the appearance of aggregates of particles within this membrane. The aggregates appear to be markers for luminal membrane osmotic water permeability. This report analyzes the relationship between transbladder osmotic water flow and aggregate frequency, and demonstrates that flow across the bladder is significantly attenuated by unstirred layers of water or by structural barriers other than the luminal membrane when the luminal membrane is made permeable by vasopressin. This analysis in addition yields unique values for the permeabilities of both the luminal membrane and the barriers to water flow which lie in series with it.

Animals↗
PubMed2002

Evidence for a central role for electro-osmosis in fluid transport by corneal endothelium.

The mechanism of transepithelial fluid transport remains unclear. The prevailing explanation is that transport of electrolytes across cell membranes results in local concentration gradients and transcellular osmosis. However, when transporting fluid, the corneal endothelium spontaneously generates a locally circulating current of approximately 25 microA cm(-2), and we report here that electrical currents (0 to +/-15 microA cm(-2)) imposed across this layer induce fluid movements linear with the currents. As the imposed currents must be approximately 98% paracellular, the direction of induced fluid movements and the rapidity with which they follow current imposition (rise time < or =3 sec) is consistent with electro-osmosis driven by sodium movement across the paracellular pathway. The value of the coupling coefficient between current and fluid movements found here (2.37 +/- 0.11 microm cm(2) hr(-1) microA (-1), suggests that: 1) the local endothelial current accounts for spontaneous transendothelial fluid transport; 2) the fluid transported becomes isotonically equilibrated. Ca(++)-free solutions or endothelial damage eliminate the coupling, pointing to the cells and particularly their intercellular junctions as a main site of electro-osmosis. The polycation polylysine, which is expected to affect surface charges, reverses the direction of current-induced fluid movements. Fluid transport is proportional to the electrical resistance of the ambient medium. Taken together, the results suggest that electro-osmosis through the intercellular junctions is the primary process in a sequence of events that results in fluid transport across this preparation.

Animals↗
PubMed2000

Water permeability measurement in living cells and complex tissues.

The identification of molecular water transporters and the generation of transgenic mice lacking water transporting proteins has created a need for accurate methods to measure water permeability. This review is focused on methodology to characterize water permeability in living cells and complex multicellular tissues. The utility of various parameters defining water transport is critically evaluated, including osmotic water permeability (P(f)), diffusional water permeability (P(d)), Arrhenius activation energies (E(a)), and solute reflection coefficients (sigma(p)). Measurements in cellular and complex tissues can be particularly challenging because of uncertainties in barrier geometry and surface area, heterogeneity in membrane transporting properties, and unstirred layer effects. Strategies to measure plasma membrane P(f) in cell layers are described involving light scattering, total internal reflection fluorescence microscopy, confocal microscopy, interferometry, spatial filtering microscopy, and volume-sensitive fluorescent indicators. Dye dilution and fluorescent indicator methods are reviewed for measurement of P(f) across cell and tissue barriers. Novel fluorescence and gravimetric methods are described to quantify microvascular and epithelial water permeabilities in intact organs, using as an example lungs from aquaporin knockout mice. Finally, new measurement strategies and applications are proposed, including high-throughput screening for identification of aquaporin inhibitors.

Animals↗