Donnan membrane equilibrium is not directly applicable to distributions of ions and water in gels or cells

Equilibration of ions and water with a charged gel does not follow the simple equations of the classical Gibbs-Donnan membrane equilibrium. Partition of ions between the gel and the external solution show specific effects, which require that activity coefficients are different in the two compartments. Highly hydrated ions, such as Na⁺ and H⁺ are accumulated into the gel water, whereas less highly hydrated ions, such as K⁺ and NH₄⁺ accumulate in the external water. This selectivity is the obverse of that found for gels containing low-density, expanded water. Water in a charged gel equilibrated with solutions of MgCl₂ was found to be more dense than bulk water at the same temperature. It is proposed that gels imbibe water to maximize the entropy of the system. Ions and water then equilibrate under those constraints. The chemical potential of water in the two compartments equalizes by an increase in density in the compartment of higher osmolality (the charged gel) and a decrease in density in the compartment of lower osmolality (the external solution). Electrolytes equilibrate so that macroscopic electroneutrality is conserved, and the chemical potential of an electrolyte is the same in each compartment. Because activity coefficients are different in the two compartments this results in asymmetric distributions of ions.Because real gels usually contain both charged and hydrophobic regions of surface, populations of water molecules of different density coexist even in very small pores. This accounts for the common failure to detect this phenomenon experimentally.     

Differences in the way potassium chloride and sucrose solutions effect osmotic potential of significance to stomata aperture modulation

Guard cell solution osmotic potential changes resulting in the opening and closing of stomata apertures follow an initial influx of potassium ions, their substitution with sucrose molecules and the subsequent reduction of the latter. To provide an insight into the osmotic mechanism of the changes, the new equation for calculating osmotic pressure, which equates the difference between the energy of pure water across a semi-permeable membrane interface with that of solution water, was used to compare the osmotic properties of KCl and sucrose. For sucrose solutions, the effect of the sucrose molecules in increasing the spacing of the solution water was mainly responsible for osmotic potential; this contrasted with K⁺ + Cl^? ions where their spacing effect was only a little higher to that of water held to those ions. At solute concentrations giving an osmotic potential level of ?3.0 MPa near that of turgid guard cells, the spacing effect on the potential of the unattached solution water molecules caused by sucrose, but in its theoretical absence, was estimated as ?2.203 MPa compared with ?1.431 MPa for KCl. In contrast, the potential attributed to water molecules firmly held to the K⁺ + Cl^? ions was ?1.212 MPa versus zero for sucrose. The potential to keep the sucrose molecules in solution was ?0.797 MPa compared with ?0.357 MPa for KCl. The findings illustrate that the way KCl effects osmotic pressure is very different to that of sucrose. It is concluded that stomata aperture modulation is closely linked to the osmotic properties of its guard cell solution solutes.     

Rectal water absorption in seawater-adapted Japanese eel Anguilla japonica

Marine teleosts drink large amounts of seawater to compensate for continuous osmotic water loss. We investigated a possible significant role of the rectum in water absorption in seawater-adapted eel. In rectal sacs filled with balanced salt solution (BSS) and incubated in isotonic BSS, water absorption was greater in seawater-adapted eel than in freshwater eel. Since rectal fluid osmolality was slightly lower than plasma osmolality in seawater-adapted eel, effects of rectal fluid osmolality on water absorption were examined in rectal sacs filled with artificial rectal fluid with different osmolality. Rectal water absorption was greater at lower rectal fluid osmolality, suggesting that an osmotic gradient between the blood and rectal fluid drives the water movement. Ouabain, a specific inhibitor of Na⁺/K⁺-ATPase, inhibited water absorption in rectal sacs, indicating that an osmotic gradient favorable to rectal water absorption was created by ion uptake driven by Na⁺/K⁺-ATPase. Expression levels of aquaporin 1 (AQP1), a water-selective channel, were significantly higher in the rectum than in the anterior and posterior intestines. Immunoreaction for Na⁺/K⁺-ATPase was detected in the mucosal epithelial cells in the rectum with more intense staining in the basal half than in the apical half, whereas AQP1 was located in the apical membrane of Na⁺/K⁺-ATPase-immunoreactive epithelial cells. The rectum is spatially separated from the posterior intestine by a valve structure and from the anus by a sphincter. Such structures allow the rectum to swell as intestinal fluid flows into it, and a concomitant increase in hydrostatic pressure may provide an additional force for rectal water absorption. Our findings indicate that the rectum contributes greatly to high efficiency of intestinal water absorption by simultaneous absorption of ions and water.     

Potassium channels in the plasmalemma ofChara corallina are multi-ion pores: Voltage-dependent blockade by Cs+ and anomalous permeabilities

Summary The outer membranes of plant cells contain channels which are highly selective for K⁺. In the giant-celled green algaChara corallina, K⁺ currents in the plasmalemma were measured when the cell was depolarized to the K⁺ equilibrium potential in relatively high external K⁺ concentrations. K⁺ current was reduced by externally added Cs⁺. Cs⁺ mainly inhibited inward K⁺ current, in a strongly voltage-dependent manner; the effective valence of the blocking reaction was often greater than 1, increasing with higher external Cs⁺ concentrations and with lower K⁺ concentrations. This is consistent with the channels being single-file, multi-ion pores. Outward current could also be inhibited by Cs⁺, when external K⁺ concentrations were low relative to Cs⁺ concentrations. As the ratio of K⁺/Tl⁺ was changed (keeping the sum of the two ions equal), both the resting potential and plasmalemma conductance went through minimums; this is the so-called anomalous mole fraction effect, and is consistent with a channel whose pore can be multiply occupied. These effects together strongly suggest that the K⁺ channels found in the plasmalemma ofChara are multi-ion pores.     

Phosphate cycles in energy crop systems with emphasis on the availability of different phosphate fractions in the soil

The growing cells of hydroponic maize roots expand at constant turgor pressure (0.48 MPa) both when grown in low-(0.5 mol m^-3 CaCl₂) or full-nutrient (Hoagland's) solution and also when seedlings are stressed osmotically (0.96 MPa mannitol). Cell osmotic pressure decreases by 0.1–0.2 MPa during expansion. Despite this, total solute influx largely matches the continuously-varying volume expansion-rate of each cell. K⁺ in the non-osmotically stressed roots is a significant exception-its concentration dropping by 50% regardless of the presence or absence of K⁺ in the nutrient medium. This corresponds to the drop in osmotic pressure. Nitrate appears to replace Cl^- in the Hoagland-grown cells.Analogous insensitivity of solute gradients to external solutes is observed in the radial distribution of water and solutes in the cortex 12 mm from the tip. Uniform turgor and osmotic pressures are accompanied by opposite gradients of K⁺ and Cl^-, outwards, and hexoses and amino acids, inwards, for plants grown in either 0.5 mol m^-3 CaCl₂ or Hoagland's solution (with negligible Cl^-). K⁺ and Cl^- levels within both gradients were slightly higher when the ions were available in the medium. The gradients themselves are independent of the direction of solute supply. In CaCl₂ solution all other nutrients must come from the stele, in Hoagland's solution inorganic solutes are available in the medium.24 h after osmotic stress, turgor pressure is recovered at all points in each gradient by osmotic adjustment using organic solutes. Remarkably, K⁺ and Cl^- levels hardly change, despite their ready availability. Hexoses are responsible for some 50% of the adjustment with mannitol for a further 30%. Some 20% of the final osmotic pressure remains to be accounted for. Proline and sucrose are not significantly involved. Under all conditions a standing water potential step of 0.2 MPa between the rhizodermis and its hydroponic medium was found. We suggest that this is due to solute leakage.Abbreviations EDX energy dispersive X-ray microanalysis - water potential - 11-1 cell osmotic pressure - P turgor pressure     

Evidence of voltage-induced channel opening in Na/K ATPase of human erythrocyte membrane

Summary Previous studies have shown that human erythrocytes when subjected to a high voltage pulsation, in the microsecond time range, lysed in an isotonic medium. The hemolysis was the result of the colloid osmotic swelling, which, in turn, was caused by the voltage perforation of the red cell membranes. In this work we demonstrate that in a low ionic medium at least 35% of the pores was related to the opening of Na⁺/K⁺ ATPase channels. The membrane conductance generated by the externally applied electric field could be partially blocked by a specific inhibitor, ouabain, or by a specific cross-linking reagent, Cu⁺⁺-phenanthroline, of the ATPase. The effect of ouabain was saturable and had a mid-point of saturation at 0.15 m. This value agrees with the physiological inhibition constant of the drug. K⁺ ion in the external medium suppressed the effect of ouabain, as has also been demonstrated in physiological studies. Experiment presented in this communication also suggests that the Na⁺/K⁺ ATPase was not perforable in a high ionic medium, and that a large fraction of the voltage-induced pores occurred at as yet unidentified sites.     

Relationship between cation accumulation and water content of salt-tolerant grasses and a sedge

Abstract Salt-tolerant grasses and a sedge were grown at three salinities in a controlled-environment greenhouse. They were measured for growth rate, ash content, water content and cations. Fourteen species from the genera Sporobolus, Aeluropus, Leptochloa, Paspalum, Puccinellia, Hordeum, Elymus, Distichlis and Spartina survived up to the highest salt treatment (540 mol m^?3 NaCl). These were designated halophytes. Eleven species from the genera Triticum, Phragmites, Dactylotenium, Cynodon, Polypogon, Panicum, Jovea and Heleocharis only survived up to 180 mol m^?3 NaCl and were designated salt-tolerant glycophytes. All species except Distichlis palmeri grew fastest on the non-saline control treatment. All species tended to have higher Na⁺ contents and lower K⁺ and water contents on saline treatments compared to control plants. Halophytes differed from glycophytes in having statistically significant lower water contents on the non-saline treatment, and lower ash contents and Na:K ratios on 180 mol m^?3. However, the range of values among species was greater than the differences between halophytes and glycophytes. All species appeared to use Na⁺ accumulation and loss of water as the main means of osmotic adjustment. Three halophytic species were grown for a longer period of time to check the above results. The osmolality of the cell sap was measured directly by the vapour pressure method and compared to calculated values based on Na⁺, K⁺ and water contents (and assuming a balancing anion such as Cl^?). Na⁺ and K⁺ alone could account for greater than 75% of the osmotic potential at all salinities. Hence, the accumulation of organic solutes did not appear to be an important factor in the osmotic adjustment of these species. The results support the conclusion that grasses coordinate Na⁺ uptake and water loss to maintain a constant osmotic potential gradient between the shoot tissues and the external solution. The results were compared to a previous study with dicotyledonous halophytes at the same location.     

Gradients in soil solution composition between bulk soil and rhizosphere – In situ measurement with changing soil water content

Soil solution composition changes with time and distance from the root surface as a result of mass flow, diffusion, plant nutrient uptake and root exudation. A model system was designed, consisting of a root compartment separated from the bulk soil compartment by a nylon net (30 m mesh size), which enabled independent measurements of the change of soil solution composition and soil water content with increasing distance from the root surface (nylon net). K⁺ concentration in the rhizosphere soil solution decreased during the initial growth stage (12 days after planting, DAP). Thereafter K⁺ accumulated with time, due to mass flow as the dominating process. The extend of K⁺ accumulation depended on the initial fertiliser application. As K⁺ concentrations in soil solution increase, not only as a result of transport exceeding uptake, but also as a result of decreasing soil water content, it is hypothesised that K concentration in soil solution is not the only trigger for the activity of K transporters in membranes, but ABA accumulation in roots induced by decreasing soil matric potentials may add to the regulation. A strong decrease of rhizosphere pH with time is observed as a result of H⁺ efflux from the roots in order to maintain cation-anion balance. In addition the K⁺ to Ca²⁺ ratio was altered continuously during the growing period, which has an impact on Ca²⁺ uptake and thus firmness of cell walls, apoplast pH, membrane integrity and activity of membrane transporters. The value of osmotic potential in the rhizosphere soil solution increased with time indicating decreasing soil water availability. Modelling approaches based on the data obtained with the system might help to fill in the time gaps caused by the low temporal resolution of soil solution sampling method.     

Responses of Atriplex spongiosa and Suaeda monoica to Salinity

The growth and tissue water, K⁺, Na⁺, Cl⁻, proline and glycinebetaine contents of the shoots and roots of two Chenopodiaceae, Atriplex spongiosa and Suaeda monoica have been measured over a range of external NaCl salinities. Both species showed some fresh weight response to low salinity mainly due to increased succulence. S. monoica showed both a greater increase in succulence (at low salinities) and tolerance of high salinities than A. spongiosa. Both species had high affinities for Na⁺ and maintained constant but low shoot K⁺ contents with increasing salinity. These trends were more marked with S. monoica in which Na⁺ stimulated the accumulation of K⁺ in roots. An association between high leaf Na⁺ accumulation, high osmotic pressure, succulence, and a positive growth response at low salinities was noted. Proline accumulation was observed in shoot tissues with suboptimal water contents. High glycinebetaine contents were found in the shoots of both species. These correlated closely with the sap osmotic pressure and it is suggested that glycinebetaine is the major cytoplasmic osmoticum (with K⁺ salts) in these species at high salinities. Na⁺ salts may be preferentially utilized as vacuolar osmotica.     

Ouabain Binding and Potassium Transport in Young and Old Populations of Human Red Cells

Human red blood cells were separated according to density by centrifugation through mixtures of phthalate esters. The densest 20% of the erythrocyte population (old cells) had reduced volume and water content compared to the lightest 20% of the cells (young cells). Corpuscular hemoglobin content was unchanged. Young cells had 50% more potassium (K⁺) than old cells, but their total intracellular concentration was only slightly higher; old cells had a small increase in sodium (Na⁺) concentration. Active K⁺ transport of young cells was 37% higher than that of old cells. [³H] + Ouabain binding revealed that this difference was the result of more K⁺ pump sites on young cells, which bound 530 ouabain molecules per cell at 100% K⁺ pump inhibition, as compared to 400 for old cells; unseparated cells bound 450-500 molecules. The relative rates of ouabain binding were identical for the two cell types. Old cells exhibited a greater passive permeability to K⁺, haying a rate coefficient for ouabain-insensitive K⁺ influx 1.8 times that of young cells. There is evidence to suggest that in the face of reduced pump activity this augmented K⁺ “leak” might enhance the osmotic stability of the old cells and function to lengthen their life span.     

Rapid Water Inflow and Outflow in Plants with Roots Treated with Salt Solutions of Various Concentrations

A new highly sensitive and rapidly responding gravimetric method was used for the investigation of rapid responses manifesting in water inflow and outflow from roots in the intact seedlings of tomato Lycopersicon esculentumMill. and sunflower Helianthus annuusL. The effects of K⁺, Na⁺, and Ca²⁺chlorides were studied in solutions with concentrations of 0.3–500 mM. Any sudden increase in the osmotic pressure of an external solution brought about a typical gravimetric response in the roots of seedlings that started practically without a lag period and comprised rapid and slow phases of water loss. The total amplitude of the response depended on the osmotic gradient produced by the changes of solutions. Responses were reversible and well reproduced upon the repeated treatment of the same plant if the treatment was noninvasive. The notable characteristics of water inflow and outflow included a very short lag period, a gradual pattern, and a low selectivity to different salts. This was especially true for the initial (rapid) phase of response. However, mono- and bivalent cations showed some specificity of action. Some data suggest the osmotic nature of rapid responses of water uptake and loss by roots. One may assume that a dynamic osmotic equilibrium exists between the root and the outer solution so that any change in the osmotic pressure of the medium would induce an instant and then a more gradual change in the water content. When plotted in logarithmic coordinates, two straight lines with different slopes described the relationship between the gravimetric response and the salt concentration. The point where these lines intersected corresponded to about 50 mM NaCl. Lower and higher salt concentrations seem to induce rapid water inflow and outflow in plant roots by means of different mechanisms.     

K+/H+ antiport in mitochondria

Mitochondria contain a latent K⁺/H⁺ antiporter that is activated by Mg²⁺-depletion and shows optimal activity in alkaline, hypotonic suspending media. This K⁺/H⁺ antiport activity appears responsible for a respiration-dependent extrusion of endogenous K⁺, for passive swelling in K⁺ acetate and other media, for a passive exchange of matrix⁴²K⁺ against external K⁺, Na⁺, or Li⁺, and for the respiration-dependent ion extrusion and osmotic contraction of mitochondria swollen passively in K⁺ nitrate. K⁺/H⁺ antiport is inhibited by quinine and by dicyclohexylcarbodiimide when this reagent is reacted with Mg²⁺-depleted mitochondria. There is good suggestive evidence that the K⁺/H⁺ antiport may serve as the endogenous K⁺-extruding device of the mitochondrion. There is also considerable experimental support for the concept that the K⁺/H⁺ antiport is regulated to prevent futile influx-efflux cycling of K⁺. However, it is not yet clear whether such regulation depends on matrix free Mg²⁺, on membrane conformational changes, or other as yet unknown factors.     

Effect of osmotic pressure,Na+/K+ ratio and medium concentration on the enzyme activity and growth of L cells in suspension culture

Summary The effect of changes in osmotic pressure and in the Na⁺/K⁺ ratio on the doubling time, maximum cell population, enzyme activity, and isoenzyme distribution pattern in suspension cultures of L cells was determined. The growth of viable cells is relatively flat over a rather wide range of osmotic pressures (220 to 440 mOsm per kg). The presence of extra salt or sucrose beyond that needed to reach the minimum osmotic pressure (220) is detrimental to cell growth as reflected by a delay in the onset of logarithmic growth, a slower growth rate, a decreased maximum population, and accelerated death phase. Excessive K⁺ ion is toxic, but the level at which it is toxic interacts with osmotic pressure of the medium. Enzyme activity and isoenzyme distribution patterns for those enzymes studied did not vary as a function of osmotic pressure, ionic ratios, or medium concentration.     

Analytical and experimental studies on the relationship between Na+, K+, and water uptake during volume increases associated with Fundulus oocyte maturation in vitro

Summary Oocytes of marine and estuarine teleosts often undergo pronounced volume increases during the maturation phase of development that precedes ovulation and fertilization. To examine the physiological correlates of these volume increases, prematuration follicles of the saltmarsh teleost, Fundulus heteroclitus, were cultured in vitro with a maturation-inducing steroid (17-hydroxy-20-dihydroprogesterone). Mean follicle volume rose significantly (75%) during a 40-h incubation period. Similar to the situation previously found in vivo, uptake of water by the maturing follicle was responsible for this volume increase in vitro, with the water content increasing from 62% to 78% of the total follicle mass. The follicle contents of two probable osmotic effectors-Na⁺ and K⁺-also rose, the increase in K⁺ being twice that of Na⁺. The influx of K⁺ even exceeded water uptake, resulting in a net increase in the concentration of this cation. It thus appears that the influx of these cations, in particular K⁺, is a major cause of the uptake of osmotically obligated water and subsequent volume increase experienced by maturing F. heteroclitus follicles. In a search for operant mechanisms, it was found that follicle hydration, but not maturation, was strictly dependent on external K⁺ in a concentration-dependent manner. The mechanism by which K⁺ accumulates in the follicle was insensitive to ouabain, so that a typical Na⁺, K⁺-ATPase mechanism does not appear to be involved. The ability of external K⁺ to promote follicle hydration was gradually lost during the maturation process as the oocyte dissociated from the surrounding granulosa cells in preparation for ovulation. Removal of all associated somatic cells prior to maturation prevented subsequent steroid-initiated hydration but not maturation. The results suggest that K⁺ may be translocated from surrounding granulosa cells to the oocyte via gap junctions during maturation.Abbreviations GVBD germinal vesicle breakdown     

Osmotic water transport with glucose in GLUT2 and SGLT

Carrier-mediated water cotransport is currently a favored explanation for water movement against an osmotic gradient. The vestibule within the central pore of Na⁺-dependent cotransporters or GLUT2 provides the necessary precondition for an osmotic mechanism, explaining this phenomenon without carriers. Simulating equilibrative glucose inflow via the narrow external orifice of GLUT2 raises vestibular tonicity relative to the external solution. Vestibular hypertonicity causes osmotic water inflow, which raises vestibular hydrostatic pressure and forces water, salt, and glucose into the outer cytosolic layer via its wide endofacial exit. Glucose uptake via GLUT2 also raises oocyte tonicity. Glucose exit from preloaded cells depletes the vestibule of glucose, making it hypotonic and thereby inducing water efflux. Inhibiting glucose exit with phloretin reestablishes vestibular hypertonicity, as it reequilibrates with the cytosolic glucose and net water inflow recommences. Simulated Na⁺-glucose cotransport demonstrates that active glucose accumulation within the vestibule generates water flows simultaneously with the onset of glucose flow and before any flow external to the transporter caused by hypertonicity in the outer cytosolic layers. The molar ratio of water/glucose flow is seen now to relate to the ratio of hydraulic and glucose permeability rather than to water storage capacity of putative water carriers.     

Fatty acid composition of lipids in vegetative organs of the halophyte Suaeda altissima under different levels of salinity

Qualitative and quantitative composition of fatty acids (FA) in the lipids of vegetative organs of the halophyte Suaeda altissima (L.) Pall. grown at different NaCl concentrations in nutrient solution was studied. Along with this, the biomass of these organs, the content of water and Na⁺, Cl^?, and K⁺ ions in them, and the ultrastructure of root and leaf cells were determined. At both low (1 mM) and high (750 mM) NaCl concentrations in nutrient solution, plants could maintain growth and water content in organs, demonstrating a noticeable increase in the dry weight and a slight increase in the water content at 250 mM NaCl. At all NaCl concentrations in nutrient solution, S. altissima tissues contained a relatively high K⁺ amount. Under salinity, Na⁺ and Cl^? ions contributed substantially into the increase in the cell osmotic pressure, i.e., a decrease in their water potential; in the absence of salinity, K⁺ fulfilled this function. In the cells of both roots and leaves, NaCl stimulated endo- and exocytosis, supposedly involved in the vesicular compound transport. 750 mM NaCl induced plasmolysis and changes in the membrane structure, which can be interpreted as degradation processes. Under optimal NaCl concentration in medium (250 mM), the content of lipids in plant aboveground organs per fresh weight was more than 2.5-fold higher than under 1 or 750 mM NaCl, whereas in the roots opposite patten was observed. When plants were grown under non-optimal conditions, substantial changes occurred in the qualitative and quantitative FA composition in lipids of both aboveground organs and roots. Observed changes are discussed in relation to processes underlying S. altissima salt tolerance and those of disintegration occurring at the high external NaCl concentration (750 mM).     

The mechanism of accumulation in living cells

When a compound enters a living cell until its activity becomes greater inside than outside, it may be said to accumulate. Since it moves from a region where its activity is relatively low to a region where its activity is relatively high, it is evident that work must be done to bring this about. The following explanation is suggested to account for accumulation. The protoplasmic surface is covered with a non-aqueous layer which is permeable to molecules but almost impermeable to ions. Hence free ions cannot enter except in very small numbers. The experiments indicate that ions combine at the outer surface with organic molecules (carrier molecules) and are thus able to enter freely. If upon reaching the aqueous protoplasm these molecules are decomposed or altered so as to set the ions free, the ions must be trapped since they cannot pass out except in very small numbers. If we adopt this point of view we can suggest answers to some important questions. Among these are the following: 1. Why accumulation is confined to electrolytes. This is evident since only ions will be trapped. 2. Why ions appear to penetrate against a gradient. Actually there is no such penetration since the ions enter in combination with molecules. The energy needed to raise the activity of entering compounds is furnished by the reactions involved in the process of accumulation. 3. Why, in absence of injury, ions do not come out when the cell is placed in distilled water. Presumably the outgoing ions will combine at the outer surface with carrier molecules and then move inward in the same way as ions coming from without. 4. Why the relative rate of penetration falls off as the external concentration increases. This is because the entrance of ions is limited by the number of carrier molecules but no such limitation exists when ions move outward since they can do so without combining with carrier molecules. 5. Why accumulation is promoted by constructive metabolism which is needed to build up the organic molecules and by destructive metabolism which brings about their decomposition. 6. Why measuring the mobilities of ions in the outer protoplasmic surface does not enable us to predict the relative rate of entrance of ions. We find for example in Nitella that K⁺ has a much higher mobility than Na⁺ but the accumulation of these ions does not differ greatly. This is to be expected if they enter by combining with molecules at the surface. Only if K⁺ is able to combine preferentially will it accumulate preferentially. 7. Why ions may come out in anoxia and at low temperatures. If these conditions depress the formation of carrier molecules and their decomposition in the protoplasm, the balance between intake and outgo of ions will be disturbed and relatively more may come out. 8. Why the excess of internal over external osmotic pressure is less in sea water than in fresh water. As the external concentration of ions increases the rate of intake does not increase in direct proportion since the number of carrier molecules does not increase and this slows down the relative rate of intake of ions. But it does not slow down the rate of exit of ions since they need not combine with carrier molecules in order to pass out. Hence the excess of ions inside will be relatively less as the concentration of external ions increases. 9. How water is pumped from solutions of higher to solutions of lower osmotic pressure. If metabolism and consequently accumulation is higher at one end of a cell than at the other, the internal osmotic pressure will be higher at the more active end and this makes it possible for the cell to pump water from solutions of higher osmotic pressure at the more active end to solutions of lower osmotic pressure at the less active, as shown experimentally for Nitella. This might help to explain the action of kidney cells and the production of root pressure in plants.     

OSMOTIC ADJUSTMENT AND DYNAMIC CHANGES IN DISTRIBUTION OF LOW MOLECULAR WEIGHT SOLUTES BETWEEN CELLULAR COMPARTMENTS OF CARROT AND BEET ROOT CELLS EXPOSED TO SALINITY

Carrot cells (Daucus carota L.) in suspension culture exposed to medium containing 150 mM NaCl plasmolyzed immediately and deplasmolyzed within 35 to 40 hr. Three days after exposure to NaCl the cells resumed proliferation. Accommodation to salinity and renewal of growth was accompanied by absorption of Na⁺ from the external medium. On completion of deplasmolysis, K⁺ concentration in the cytosol doubled and Na⁺ concentration approximated that of K⁺. The vacuolar K⁺ concentration was practically unchanged while Na⁺ accumulated to a concentration double that of K⁺. Cl^−- accumulation started later and eventually exceeded that of Na⁺ plus K⁺. Malate was redistributed during accommodation to salinity and eventually returned to its initial level. Amino acid content in the cytosol increased fivefold, while in the vacuole it remained unchanged. These results show that: 1) recovery from osmotic shock requires absorption of easily penetrating solute, mainly Na⁺; 2) distribution of solutes, absorbed or synthesized in cells exposed to salinity, is a dynamic process; 3) cells could grow and proliferate in high NaCl content in the cytosol; 4) red beet root cells grown in the presence of NaCl contain higher cytoplasmic Na⁺ than K⁺; and 5) during adjustment to salinity small spherical carrot cells survive the osmotic shock and do not show any detectable damage.     

Solute Transport Process in Intestinal Epithelial Cells

In rat small intestine, the active transport of organic solutes results in significant depolarization of the membrane potential measured in an epithelial cell with respect to a grounded mucosal solution and in an increase in the transepithelial potential difference. According to the analysis with an equivalent circuit model for the epithelium, the changes in emf's of mucosal and serosal membranes induced by active solute transport were calculated using the measured conductive parameters. The result indicates that the mucosal cell membrane depolarizes while the serosal cell membrane remarkably hyperpolarizes on the active solute transport. Corresponding results are derived from the calculations of emf's in a variety of intestines, using the data that have hitherto been reported. The hyperpolarization of serosal membrane induced by the active solute transport might be ascribed to activation of the serosal electrogenic sodium pump. In an attempt to determine the causative factors in mucosal membrane depolarization during active solute transport, cell water contents and ion concentrations were measured. The cell water content remarkably increased and, at the same time, intracellular monovalent ion concentrations significantly decreased with glucose transport. Net gain of glucose within the cell was estimated from the restraint of osmotic balance between intracellular and extracellular fluids. In contrast to the apparent decreases in intracellular Na⁺ and K⁺ concentrations, significant gains of Na⁺ and K⁺ occurred with glucose transport. The quantitative relationships among net gains of Na⁺, K⁺ and glucose during active glucose transport suggest that the coupling ratio between glucose and Na⁺ entry by the carrier mechanism on the mucosal membrane is approximately 1:1 and the coupling ratio between Na⁺-efflux and K⁺-influx of the serosal electrogenic sodium pump is approximately 4:3 in rat small intestine. In addition to the electrogenic ternary complex inflow across the mucosal cell membrane, the decreases in intracellular monovalent ion concentrations, the temporary formation of an osmotic pressure gradient across the cell membrane and the streaming potential induced by water inflow through negatively charged pores of the cell membrane in the course of an active solute transport in intestinal epithelial cells are apparently all possible causes of mucosal membrane depolarization.     

Coupling of water and potassium ions in K+ channels of the tonoplast of Chara

Electrokinetic measurements, of streaming potential, were carried out on an excised inside-out patch of the vacuolar membrane of Chara corallina. A water activity gradient was imposed across the patch membrane containing a single K⁺ channel by addition of sorbitol to one side. Two different K⁺ channels were found in the tonoplast. Their open channel conductance was investigated as a function of KCl concentration. They had a maximal open channel conductance of 247 and 173 pS, and an apparent affinity (K_M) of 116 and 92 mM, respectively. Single-channel zero-current potentials were determined in the presence of an osmotic gradient, and dilution artifacts were corrected for by addition of valinomycin to the bath. Our results suggest that 29 water molecules were coupled to the transport of one K⁺ ion in the large conductance K⁺ channel which has a pore radius of ~1.5 nm.