Osmotic water flow in leaky epithelia

I review three currently unsolved and controversial problems in understanding solute-linked water transport in epithelia. 1. Values of osmotic water permeability (Posm) calculated from steady-state osmotic flow in response to a gradient of a probe molecule tend to be underestimates, because of three unstirred-layer (USL) effects. These are: dissipation of the probe's gradient by diffusion in USL's; reduction of the probe's gradient, due to the sweeping-away effect of water flow generated by the probe itself; and solute polarization (creation of an opposing gradient of an initially symmetrically distributed solute by the sweeping-away effect). These errors increase with probe permeability, USL thickness, Posm, and concentration ratio of symmetrically distributed solute to probe, and vary inversely as the fractional area available for water flow (e.g., lateral intercellular space width). The form of an osmotic transient, and the possibility of extracting a true Posm value from the transient, depend on the relative values of three time constants: those for solute diffusion in USL's, for solute polarization by water flow in USL's and for measuring water flow. Sweeping-away effects cause major underestimates (by one or more orders of magnitude) in epithelial Posm determinations, as shown by apparent streaming potentials during osmotic flow and by transiently reversed flows after removal of the proble. True Posm values for leaky epithelia probably exceed 10(-3) or 10(-2) cm/sec.osm. The necessary conditions for resolving osmotic transients are set out. 2. I illustrate the difficulties in deciding what fraction of transepithelial water flow is via the cells, and what fraction via the junctions. There is no existing method for answering this question. 3. Controversies about the validity, or need for modification, of the standing-gradient theory are discussed. Progress in this field requires new methods: to resolve osmotic transients; to separate transcellular and transjunctional water flows; and to measure solute concentrations in lateral intercellular spaces directly.     

Volume flows across gallbladder epithelium induced by small hydrostatic and osmotic gradients

Summary The hydraulic conductivity of rabbit gallbladder epithelium has been studied using a continuous volumetric method based on capacitance measurements. The time resolution for measuring osmotic flows is in the range of seconds. Volume flows have been induced by osmotic gradients between 0 and 100 mosmol. In this range the flow-force relation is linear and theP _f value is 9.3×10^–3 cm/sec. After correction for solute polarization effects, theP _f value amounts to 0.05 cm/sec. The observed flow is constant between 5 sec up to 20 min after a sudden increase in the osmolarity of the mucosal solution. The wet weight of the gallbladder tissue decreases by 22% and increases by 30% during osmotic flows from serosa to mucosa and from mucosa to serosa, respectively. Volume flows induced by hydrostatic pressure gradients on the mucosal surface are linearly related to the driving forces between 0 and 40 mbar. TheP _f value is 0.15 cm/sec. The volume flows are constant between 2 sec and 15 min after pressure application. The flow-force relation for pressure gradients on the serosal surface is markedly nonlinear for gradients greater than 5 mbar. Below 5 mbar theP _f value is 4.5 cm/sec. From electrical measurements, e.g., resistance and streaming potentials, and from flux studies with inulin and polyethylene glycol 4000, it is concluded that hydrostatic and osmotic gradients are not comparable when they are applied to gallbladder epithelium. They induce volume flows across different pathways, e.g., osmosis predominantly across the cellular route and pressure filtration predominantly across paracellular routes.     

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.     

The development of osmotic flow through an unstirred layer

We investigate the errors involved in estimating the osmotic permeability of a semi-permeable membrane, from the measured osmotic flow and the difference in concentration of osmotically active solute across it, without taking account of the unstirred layer in the solution next to the membrane. In the problem solved, this layer is represented as a region of thickness δ at the far side of which a solute concentration C_b is imposed for time . The initial diffusion of solute towards the membrane causes the concentration at the membrane C_m to rise, generating an osmotic flow of water, J, whose convective effect opposes the diffusion. The problem is made non-linear by the dependence of J on C_m. Ultimately a steady state is set up, in which C_m is less than C_b. The solution is shown to depend on a single parameter β, equal to (L_pRT) δ C_b/D, where L_pRT is the osmotic permeability of the membrane and D is the diffusivity of solute. Solution of the steady state leads to a prediction of C_m/C_b as a function of β, and analysis of the decay of transient terms leads to a prediction of the decay time π, also as a function of β. Numerical data for membranes with a wide range of osmotic permeabilites, and for a reasonable range of solute, i.e. sucrose, concentrations, suggest that values of β can range from 0.001 or below to 7.5 or above. The former value implies negligible error in neglecting the unstirred layer, while the latter implies a 79%. error. For β = 0.1 and for δ = 2 × 10⁻⁴ m, π is predicted to be around 74 s. This decreases as β increases (for fixed δ); for values of β above about 27, the decay of transients is no longer monotonic but takes the form of damped oscillations.     

Effects of electrical gradients on volume flows across gall bladder epithelium

A volumetric method has been developed which permits continuous registration of volume flows across epithelial tissues. The method was applied to volume flow measurements across rabbit gall bladder epithelium. The rate of fluid reabsorption measured in this way was twice as high as previously observed in sac preparations of the gall bladder. This is probably due to better aeration and stirring of the mucosal solution. It was demonstrated that electrical gradients across the gall bladder induced volume flows towards the negative electrode. In non-transporting bladders volume flows were linearly related with current between 300 and 900 μA in both directions. However, volume flow rates were three times higher from mucosa to serosa than in the opposite direction. From the magnitude of polarization potentials, observed after switching off the current, the conclusion was reached that all of the current-induced volume flow is an osmotic flow due to salt polarization in the unstirred layers of the tissue. By implication, so-called streaming potentials observed during osmotic flows reflect solely polarization effects. In actively transporting gall bladders a 200 μA current increased or decreased the flow rate twice as much as expected from polarization effects alone. Therefore passage of current interfered directly with the active transport mechanism of gall bladder epithelium.     

OZONE-INDUCED MEMBRANE PERMEABILITY CHANGES

The effect of 0.5 ppm ozone for 0.5-1 hr on plant cell membrane permeability was ascertained. Permeabilities to both water and solutes were estimated by measuring leaf disc weight changes and following tritiated water and ⁸⁶Rb fluxes. Measurements were made immediately after ozone exposure and 24 hr after exposure. The reflection coefficient, σ, an index of solute permeability, decreased in ozone-treated primary leaves of pinto bean (Phaseolus vulgaris). The latter indicates an increase in membrane solute permeability or internal solute leakage. Water and THO flux estimates both indicated a decrease in membrane permeability to water; both the hydraulic conductivity (L_p) and the water diffusional coefficient (L_D) apparently decreased, an anomaly which is discussed. These data indicate that ozone has a direct effect on membrane function by altering permeability characteristics. We assume from these data that cell membranes are primary target sites for ozone injury.     

Characteristics of Hg- and Zn-sensitive Water Channels in the Plasma Membrane of Chara Cells*

Hydraulic conductivity (L_p) of the plasma membrane of Chara corallina was inhibited by HgCl₂ maximally by about 95%. The inhibition was reversed by 2-mercaptoethanol, reconfirming the observation obtained by Henzler and Steudle (1995). The results suggest that osmotic water transport through Chara cells occurs mostly via mercury-sensitive water channels containing thiol groups. ZnCl₂ dissolved in APW (pH 5.6) also inhibited L_p by about 80% within 1–2 h, while ZnCl₂ dissolved in Hepes-Tris buffer (pH 7.4) inhibited it by about 90% within several minutes. Inhibition of L_p by ZnCl₂ was also reversed by 2-mercaptoethanol, suggesting that zinc acts also on thiol groups of water channel proteins. Cells from which tonoplast had been removed by ECTA were as sensitive to both HgCl₂ and ZnCl₂ (pH 7.4) as normal cells. This demonstrates that water channels sensitive to thiol reagents really exist in the plasma membrane. On the other hand, ZnCl₂ (pH 5.6) did not inhibit L_p of tonoplast-free cells. This may be accounted for by assuming first that Hg- and Zn-sensitive thiol groups of water channels may exist on the cytoplasmic side, and second that ZnCl₂ in acidic medium may exist in ionized species which can be chelated by EGTA after permeation. The polar water permeability, or the endoosmotic L_p being larger than the exoosmotic one, was not affected by lowering the rate of osmosis by decreasing the osmotic gradient for transcellular osmosis down to 0.02 M sorbitol. The polarity disappeared when osmotic water flow through water channels was completely inhibited by HgCl₂. Thus the polarity is assumed to be intrinsic to water channels in the plasma membrane.     

The role of membrane in osmotic flow

Osmosis as a phenomenon caused by internal forces goes on without the necessity for the presence of any external forces. Therefore pressure gradient plays no special role in osmotic flow. Membrane as a component of solution with its molecules possessing some kind of mobility ceases to be a passive obstacle to the flow of other components, but becomes also a co-determining factor in osmotic flow. This has been shown by using the methods of irreversible thermodynamics. In the state of osmotic equilibrium osmosis does not occur. So also the mobility of water molecules which may then be found in tracer experiment does not determine the osmotic permeability coefficient. The coefficientsσ andω as defined by the parameters of the system under the condition of zero volume flow are not directly connected to L_p.     

Pseudo-streaming potentials in Necturus gallbladder epithelium. II. The mechanism is a junctional diffusion potential

The mechanisms of apparent streaming potentials elicited across Necturus gallbladder epithelium by addition or removal of sucrose from the apical bathing solution were studied by assessing the time courses of: (a) the change in transepithelial voltage (Vms). (b) the change in osmolality at the cell surface (estimated with a tetrabutylammonium [TBA+]-selective microelectrode, using TBA+ as a tracer for sucrose), and (c) the change in cell impermeant solute concentration ([TMA+]i, measured with an intracellular double-barrel TMA(+)-selective microelectrode after loading the cells with TMA+ by transient permeabilization with nystatin). For both sucrose addition and removal, the time courses of Vms were the same as the time courses of the voltage signals produced by [TMA+]i, while the time courses of the voltage signals produced by [TBA+]o were much faster. These results suggest that the apparent streaming potentials are caused by changes of [NaCl] in the lateral intercellular spaces, whose time course reflects the changes in cell water volume (and osmolality) elicited by the alterations in apical solution osmolality. Changes in cell osmolality are slow relative to those of the apical solution osmolality, whereas lateral space osmolality follows cell osmolality rapidly, due to the large surface area of lateral membranes and the small volume of the spaces. Analysis of a simple mathematical model of the epithelium yields an apical membrane Lp in good agreement with previous measurements and suggests that elevations of the apical solution osmolality elicit rapid reductions in junctional ionic selectivity, also in good agreement with experimental determinations. Elevations in apical solution [NaCl] cause biphasic transepithelial voltage changes: a rapid negative Vms change of similar time course to that of a Na+/TBA+ bi-ionic potential and a slow positive Vms change of similar time course to that of the sucrose-induced apparent streaming potential. We conclude that the Vms changes elicited by addition of impermeant solute to the apical bathing solution are pseudo-streaming potentials, i.e., junctional diffusion potentials caused by salt concentration changes in the lateral intercellular spaces secondary to osmotic water flow from the cells to the apical bathing solution and from the lateral intercellular spaces to the cells. Our results do not support the notion of junctional solute-solvent coupling during transepithelial osmotic water flow.     

Electrolytes control flows of water across the apical barrier in toad skin: The hydrosmotic salt effect

Summary The reversible dependence of skin osmotic water permeability (L _PD ) upon the ionic concentration of the outer bathing solution — which we have called hydrosmotic salt effect (HSE) — was studied in the isolated skin of the toadBufo marinus ictericus. The skin osmotic water flow (J _V ) was measured as a function of outer bathing solution osmolality (O _e ).L _PD , calculated as (J _v /) _P=0 (where and P are the osmotic and hydrostatic pressure differences across the skin, respectively) was constant whenO _e was altered with sucrose, a nonelectrolyte. In contrast,L _PD increased continuously in the hypotonic range asO _e was raised from zero (distilled water) with NaCl or KCl. The HSE could also be evoked in the condition of reversed osmotic volume flow, with the outer bathing medium made hypertonic with sucrose.Diffusional¹⁴C-sucrose permeability, measured in theJ _v =0 condition to prevent solvent drag of sucrose in the paracellular pathways, indicate that the hydrosmotic salt effect cannot be explained by assuming a paracellular permeability increase, due to tight junction opening, but might be interpreted as due to changes in the osmotic water permeability of the apical membranes of the most superficial cells of the epithelium.The hydrosmotic salt effect can be elicited in control skins and in vasopressin-stimulated skins, on top of the hormonal response.The time course of the hydrosmotic salt effect is substantially different from that of the hydrosmotic response to vasopressin. Its half-time is 4 to 5 times faster than that of vasopressin action, with individual values as short as 1.5 min.The time courses of the hydrosmotic salt-effect onset and reversibility are exponential, clearly contrasting with the typical sigmoidal shape of vasopressin onset and washout time courses.Based on time course data and on speed of response we postulate that the mechanism underlying the hydrosmotic salt effect is due to modifications of existing water pathways in the apical membrane, rather than to incorporation and removal of water permeability units in this structure.     

Unstirred layer effects in osmotic water flow across gallbladder epithelium

Summary The standard one-dimensional model of the unstirred layer is applied in a re-examination of the experimental results of Wright, Smulders and Tormey (Wright, E.M., Smulders, A.P., Tormey, J. McD., 1972,J. Membrane Biol. 7:198) who reported large transients in the osmotic flux of water from the serosal to the mucosal side of rabbit gallbladder epithelium. They initiated osmosis by the addition of sucrose to the mucosal bathing solution (initially, approximately 300mOsm NaCl) and observed that the initial flux was more than ten times its eventual steady-state value; they interpreted this as a consequence of the piling-up of NaCl in the unstirred tissue layer on the serosal side of the epithelium. The present analysis (both steady-state and unsteady) shows that if measured values of layer thickness are used, together with reasonable values of the reduced diffusivity of NaCl in the tissue and of the fraction of tissue available for water flow, then one would predict a discrepancy of only about 10%, not tenfold, between the initial and final values of the flux. Thus the standard model is inconsistent with the observations. Furthermore, Wright et al's results cannot be used to infer that the osmotic permeability of epithelial cell membranes is much larger than steadystate measurements on whole epithelia would indicate. Mucosal-to-serosal flow is also analyzed, and in this case a considerably greater osmotic permeability is predicted; this result is consistent with the observed changes in structure of the lateral intercellular spaces when the direction of flow is reversed.     

Membrane Permeability Modeling: Kedem–Katchalsky vs a Two-Parameter Formalism

The analysis of experiments for the purpose of determining cell membrane permeability parameters is often done using the Kedem–Katchalsky (KK) formalism (1958). In this formalism, three parameters, the hydraulic conductivity (L_p), the solute permeability (P_s), and a reflection coefficient (ς), are used to characterize the membrane. Sigma was introduced to characterize flux interactions when water and solute (cryoprotectant) cross the membrane through a common channel. However, the recent discovery and characterization of water channels (aquaporins) in biological membranes reveals that aquaporins are highly selective for water and do not typically cotransport cryoprotectants. In this circumstance, sigma is a superfluous parameter, as pointed out by Kedem and Katchalsky. When sigma is unneeded, a two-parameter model (2P) utilizing onlyL_pandP_sis sufficient, simpler to implement, and less prone to spurious results. In this paper we demonstrate that the 2P and KK formalism yield essentially the same result (L_pandP_s) when cotransporting channels are absent. This demonstration is accomplished using simulation techniques to compare the transport response of a model cell using a KK or 2P formalism. Sigma is often misunderstood, even when its use is appropriate. It is discussed extensively here and several simulations are used to illustrate and clarify its meaning. We also discuss the phenomenological nature of transport parameters in many experiments, especially when both bilayer and channel transport are present.     

Effect of osmolality on the hydraulic permeability coefficient of red cells

The osmotic water permeability coefficient, L_p, for human and dog red cells has been measured as a function of medium osmolality, and found to depend on the osmolality of the bathing medium. In the case of human red cells L_p falls from 1.87 x 10^-11 cm³/dyne sec at 199 mOSM to 0.76 x 10^-11 cm³/dyne sec at 516 mOSM. A similar decrease was observed for dog red cells. Moreover, L_p was independent of the direction of water movement and the nature of the solute used to provide the osmotic pressure gradient; it depended only on the final osmolality of the medium. Furthermore, L_p was not affected by pH in the range of 6 to 8 nor by the presence of drugs such as valinomycin (1 x 10^-6 M) and tetrodotoxin (3.2 x 10^-6 M). The instantaneous nature of the response to changes in external osmolality suggests that the hydraulic conductivity of the membrane is controlled by a thin layer at the outer face of the membrane.     

The effect of osmotically induced water flows on the permeability and ultrastructure of the rabbit gallbladder

Summary The purpose of these experiments was to determine the effect of osmotic gradients on the permeability of the rabbit gallbladder. Increasing the tonicity of the mucosal solution reduced the permeability of the gallbladder to both ions and nonelectrolytes, whereas there was no significant effect when the serosal solution was made hypertonic. These results cannot be explained by solvent/solute interactions in either the epithelial membranes or the unstirred layers. Associated with the changes in permeability was the appearance of the transport number effect and current induced resistance changes. Morphological studies of the gallbladder under these conditions showed that the extracellular spaces of the epithelium and the rest of the wall dilate in the presence of osmotic flow to the serosa, but that the spaces collapse when the flow is in the opposite direction. Reconstruction of the permeability changes from the dimensions of the tissue show that all the physiological phenomena are accounted for by the changes in morphology, the dominant effect being in the lateral intercellular spaces. These results suggest that the lateral spaces are a common pathway shared by all solutes crossing the epithelium, and that diffusion along these spaces becomes rate limiting as the spaces collapse.     

Concentration Polarization in an Ultrafiltering Capillary

Concentration polarization, the accumulation of retained solute next to an ultrafiltering membrane, elevates osmotic pressure above that which would exist in the absence of polarization. For ultrafiltration in a cylindrical tube, use of the radially averaged solute concentration results in an underestimate of osmotic pressure, yielding an effective hydraulic permeability (k) less than the actual membrane hydraulic permeability (k_m). The extent to which k and k_m might differ in an ultrafiltering capillary has been examined theoretically by solution of the momentum and species transport equations for idealized capillaries with and without erythrocytes. For diameters, flow velocities, protein concentrations and diffusivities, and ultrafiltration pressures representative of the rat glomerular capillary network, results indicate that the effects of polarization are substantial without erythrocytes (k/k_m = 0.7) and persist, but to a lesser extent, with erythrocytes (k/k_m = 0.9), the reduction in polarization in the latter case being due to enhanced plasma mixing. In accord with recent experimental findings in rats, k is found to be relatively insensitive to changes in glomerular plasma flow rate.     

Amicus Plato, sed magis amica veritas: plots must obey the laws they refer to and models shall describe biophysical reality!

In the companion paper, we discussed in details proper linearization, calculation of the inactive osmotic volume, and analysis of the results on the Boyle-vant’ Hoff plots. In this Letter, we briefly address some common errors and misconceptions in osmotic modeling and propose some approaches, namely: (1) inapplicability of the Kedem–Katchalsky formalism model in regards to the cryobiophysical reality, (2) calculation of the membrane hydraulic conductivity L_p in the presence of permeable solutes, (3) proper linearization of the Arrhenius plots for the solute membrane permeability, (4) erroneous use of the term “toxicity” for the cryoprotective agents, and (5) advantages of the relativistic permeability approach (RP) developed by us vs. traditional (“classic”) 2-parameter model.     

Osmotic relations of the drought-tolerant shrub Artemisia tridentata in response to water stress

Turgor maintenance, solute content and recovery from water stress were examined in the drought-tolerant shrub Artemisia tridentata. Predawn water potentials of shrubs receiving supplemental water remained above ?2 MPa throughout summer, while predawn water potentials of untreated shrubs decreased to ?5 MPa. Osmotic potentials decreased in conjunction with water potentials maintaining turgor pressures above 0 MPa. The decreases in osmotic potentials were not the result of osmotic adjustment (i.e. solute accumulation). Leaf solute contents decreased during drought, but leaf water volumes decreased more than 75% from spring to summer, thereby passively concentrating solutes within the leaves. The maintenance of positive turgor pressures despite decreases in leaf water volumes is consistent with other studies of species with elastic cell walls. Inorganic ion, organic acid, and carbohydrate contents of leaves declined during drought. The only solutes accumulating in leaves of A. tridentata with water stress were proline and a cyclitol, both considered compatible solutes. Total and osmotic potentials recovered rapidly following rewatering of shrubs; solute contents did not change except for a decrease in proline. Maintaining turgor through the passive concentration of solutes may be advantageous compared to synthesis of new solutes for osmotic adjustment in arid environments.     

Barrier components acting against osmotic water flow inNitella

In order to check whether or not the layer of chloroplasts densely arranged in the cortical gel of aNitella internode offers substantial resistance to osmotic water flow, a material was prepared which had the cortical gel layer freed from chloroplasts by centrifugation either longitudinal or lateral to the cell axis. The water permeability of the cell remained the same as normal even though the chloroplasts were exfoliated from the cell cortex to the extent of 50% of the total area, showing that the chloroplast layer plays hardly any significant part as a barrier to osmotic flow. Since it is known that the layer of the streaming endoplasm is also negligible as resistance against osmotic water flow (Tasawa and Kamiya, 1965), it is concluded that the major barrier components against osmotic flow in theNitella internode are the cell wall, plasmalemma and/or tonoplast.     

Contribution of solvent drag through intercellular junctions to absorption of nutrients by the small intestine of the rat

Summary The lumen of the small intestine in anesthetized rats was recirculated with 50 ml perfusion fluid containing normal salts, 25mm glucose and low concentrations of hydrophilic solutes ranging in size from creatinine (mol wt 113) to Inulin (mol wt 5500). Ferrocyanide, a nontoxic, quadrupally charged anion was not absorbed; it could therefore be used as an osmotically active solute with reflection coefficient of 1.0 to adjust rates of fluid absorption,J _v , and to measure the coefficient of osmotic flow,L _p . The clearances from the perfusion fluid of all other test solutes were approximately proportional toJ _v . FromL _p and rates of clearances as a function ofJ _v and molecular size we estimate (a) the fraction of fluid absorption which passes paracellularly (approx. 50%), (b) coefficients of solvent drag of various solutes within intercellular junctions, (c) the equivalent pore radius of intercellular junctions (50 Å) and their cross sectional area per unit path length (4.3 cm per cm length of intestine). Glucose absorption also varied as a function ofJ _v . From this relationship and the clearances of inert markers we calculate the rate of active transport of glucose, the amount of glucose carried paracellularly by solvent drag or back-diffusion at any givenJ _v and luminal glucose concentration and the concentration of glucose in the absorbate. The results indicate that solvent drag through paracellular channels is the principal route for intestinal transport of glucose or amino acids at physiological rates of fluid absorption and concentration. In the absence of luminal glucose the rate of fluid absorption and the clearances of all inert hydrophilic solutes were greatly reduced. It is proposed that Na-coupled transport of organic solutes from lumen to intercellular spaces provides the principal osmotic force for fluid absorption and triggers widening of intercellular junctions, thus promoting bulk absorption of nutrients by solvent drag. Further evidence for regulation of channel width is provided in accompanying papers on changes in electrical impedance and ultrastructure of junctions during Na-coupled solute transport.