Potassium transport in normal and transformed mouse 3T3 cells

The components of unidirectional K influx and efflux have been investigated in the 3T3 cell and the SV40 transformed 3T3 cell in exponential and stationary growth phase. Over the cell densities used for transport experiments the 3T3 cell goes from exponential growth to density dependent inhibition of growth (4 × 10⁴ to 4 × 10⁵ cell cm^?2) whereas the SV40 3T3 maintains exponential or near exponential growth (4 × 10⁴ to 1 × 10⁶ cell cm^?2). In agreement with previous observations, volume per cell and mg protein per cell decrease with increasing cell density. Thus, transport measurements have been expressed on a per volume basis. Total unidirectional K influx and efflux in the 3T3 cell is approximately double that of the SV40 3T3 cell at all cell densities investigated. Both cell types have similar volumes initially and show similar decreases with increasing cell density. Thus, in this clone of the 3T3 cell SV40 transformation specifically decreases unidirectional K flux. The magnitude of the total K flux does not change substantially for either cell line during transition from sparse to dense cultures. However, the components of the K transport undergo distinct changes. Both cell lines possess a ouabain sensitive component of K influx, presumably representing the active inward K pump. Both also possess components of K influx and efflux sensitive to furosemide. The data suggest this component represents a one-for-one K exchange mechanism. The fraction of K influx mediated by the ouabain sensitive component is reduced to one half its value when exponential versus density inhibited 3T3 cells are compared (63% versus 31% of total influx). No comparable drop occurs in the SV40 3T3 cell at equivalent cell densities (64% versus 56% of total influx). Thus, the pump mediated component of K influx would appear to be correlated with growth. In contrast, the furosemide sensitive component represents approximately 20% of the total unidirectional K influx and efflux in both cell lines in sparse culture. At high cell densities, where growth inhibition occurs in the 3T3 cell but not the SV40 3T3, the furosemide sensitive component doubles in both cell lines. Thus, the apparent K-K exchange mechanism is density dependent rather than growth dependent.     

Potassium fluxes in dialyzed squid axons

Measurements have been made of K influx in squid giant axons under internal solute control by dialysis. With [ATP]_i = 1 µM, [Na]_i = 0, K influx was 6 ± 0.6 pmole/cm² sec; an increase to [ATP]_i = 4 mM gave an influx of 8 ± 0.5 pmole/cm² sec, while [ATP]_i 4, [Na]_i 80 gave a K influx of 19 ± 0.7 pmole/cm² sec (all measurements at ∼16°C). Strophanthidin (10 µM) in seawater quantitatively abolished the ATP-dependent increase in K influx. The concentration dependence of ATP-dependent K influx on [ATP]_i, [Na]_i, and [K]_o was measured; an [ATP]_i of 30 µM gave a K influx about half that at physiological concentrations (2–3 mM). About 7 mM [Na]_i yielded half the K influx found at 80 mM [Na]_i. The ATP-dependent K influx responded linearly to [K]_o from 1–20 mM and was independent of whether Na, Li, or choline was the principal cation of seawater. Substances tested as possible energy sources for the K pump were acetyl phosphate, phosphoarginine, PEP, and d-ATP. None was effective except d-ATP and this substance gave 70% of the maximal flux only when phosphoarginine or PEP was also present.     

Potassium Fluxes in Desheathed Frog Sciatic Nerve

Desheathed frog (R. pipiens) sciatic nerves were soaked in Na-deficient solutions, and measurements were made of their Na and K contents and of the movements of K⁴². When a nerve is in Ringer's solution, the Na fluxes are equal to the K fluxes, and about 75 per cent of the K influx is due to active transport. The Na content and the Na efflux are linearly related to the Na concentration of the bathing solution, while the K content and the K fluxes are not so related. When a nerve is in a solution in which 75 per cent of the NaCl has been replaced by choline chloride or sucrose, the active K influx exceeds the active Na efflux, and the K content is maintained. When a nerve is soaked in a solution that contains Li, the K⁴² uptake is inhibited, and the nerve loses K and gains Li. When a Li-loaded nerve recovers in a Li-free solution, K is taken up in exchange for Li. This uptake of K requires Na in the external solution. It is concluded that the active transports of K and of Na may be due to different processes, that an accumulation of K occurs only in exchange for an intracellular cation, which need not be Na, and that Na plays a specific, but unknown, role in K transport.     

Acclimation of potassium influx in rye (Secale cereale) to low root temperatures

The influx of K⁺(⁸⁶Rb⁺) into intact roots of rye (Secale cereale L. cv. Rheidal) exposed to a differential temperature (DT) between the root (8° C) and shoot (20° C) is initially reduced compared with warm-grown (WG) controls with both shoot and root maintained at 20° C. Over a period of 3 d, however, K⁺-influx rates into DT plants are restored to levels similar to or greater than those of the WG controls, the absolute rates of K⁺ influx being strongly dependent upon the shoot/root ratio. Acclimation in DT plants results in a reduction of K⁺ influx into the apical (0–2 cm) region of the seminal root which is associated with a compensatory increase in K⁺ influx into the more mature, basal regions of the root. Values of V _max and apparent K _m for K⁺ influx into DT plants were similar to those for WG plants at assay temperatures of 8° C and 20° C except for an increase in the apparent K _m at 8° C. The influx of K⁺ from solutions containing 0.6 mol·m^-3 K⁺ into both WG and DT plants was found to be linearly related to assay temperature over the range 2–27° C, and the temperature sensitivity of K⁺ influx to be dependent upon shoot/root ratio. At high shoot/root ratios, the ratio of K⁺ influx at 20° C:K⁺ influx at 8° C for WG plants approached a minimum value of 1.9 whereas that for DT plants approached unity indicating that K⁺ influx into DT plants has a large temperature-insensitive component. Additionally, when plants were grown in solutions of low potassium concentration, K⁺ influx into DT plants was consistently greater than that into WG plants, in spite of having a greater root potassium concentration ([K⁺]int). This result indicates some change in the regulation of K⁺ influx by [K⁺]int in plants exposed to low root temperatures. We suggest that K⁺ influx into rye seedlings exposed to low root temperatures is regulated by the increased demand placed on the root system by a proportionally larger shoot and that the acclimation of K⁺ influx to low temperatures may be the result of an increased hydraulic conductivity of the root system.Abbreviations DT differential temperature pretreatment - [K⁺]int root potassium concentration - [K⁺]ext potassium concentration of nutrient medium - WG warm-grown pretreatment     

Cat Heart Muscle in Vitro : VI. Potassium exchange in papillary muscles

The exchange of cell K with K⁴², J _K, has been measured in cat right ventricular papillary muscle under conditions of a steady state with respect to intracellular K concentration. Within the limits of the measurement, all of cell K exchanged at a single rate. Cells from small cats are smaller and have larger surface/volume ratios than cells from large cats. The larger surface/volume ratio results in larger flux values. J _K increases in an approximately linear manner as the external K concentration is increased twentyfold, from 2.5 to 50 mM, at constant intracellular K concentration. The permeability for K ions, P _K, calculated from the influx and membrane potential, remains very nearly constant over this range of external K concentrations. J _K is not affected by replacement of O₂ by N₂, or by stimulated contractions at 60 per minute, but K influx decreases markedly in 10^-5 M and 10^-8 M ouabain.     

Effects of Leaf Age and Position on the Shoot Axis on Potassium Absorption by Tomato Leaf Slices

The effect of leaf age on K (⁸⁶Rb) influx into tomato (Lycopersiconesculentum Mill.) leaf lamina slices was determined for leaves5, 9 and 13 counting acropetally. Potassium influx rates expressedon a leaf fresh weight basis declined rapidly during leaf elongationat external KCI concentrations between 0.5 and 20.0 mM. In fullyexpanded leaves, K influx rates declined more slowly with age.The onset of senescence in mature leaves did not result in alarge loss in K uptake capability. Leaf position on the shootaxis and the stage of whole plant development had little influenceon K influx into leaf cells. It is suggested that the rapiddecrease in K influx in growing leaves is related to a dilutionin the concentration of K transporter sites resulting from anincrease in cell volume and weight. Lycopersicon esculentum Mill, tomato, free space, potassium, influx rate, ion uptake, leaf slices, leaf age leaf ontogeny     

Potassium transport and content during G1 and S phase following serum stimulation of 3T3 cells.

The effect of serum stimulation on unidirectional and net K flux and their relationship to the initiation of DNA synthesis has been investigated in mouse 3T3 fibroblasts. Stimulation of quiescent 3T3 cells with 20% serum results in the initiation of S phase approximately ten hours after serum addition. During transition from G₁ to S phase distinct changes in K transport and cellular K content occur. Total unidirectional K influx undergoes an immediate 2-fold increase upon serum addition, an observation in qualitative agreement with previous results (Rozengurt and Heppel, 1975). This total increase in unidirectional K influx represents a proportional increase in the active, ouabain sensitive component and the K-K exchange component. The initial increase in total flux is followed by a gradual decline over a 16-hour period to levels approaching those of quiescent cells. Following the initial increase in unidirectional K influx is an approximately 75% increase in cell K on a per milligram protein basis or a 40% increase on a per volume basis. This increase peaks at four to five hours and then declines to initial levels at 10 to 14 hours. Populations of quiescent cells given 20% serum plus 0.5 mM ouabain simultaneously are totally blocked from entering S phase, as determined by the appearance of ³H-thymidine labeled nuclei. However, if the ouabain is removed after six hours these cells then undergo the same changes in unidirectional K influx and content as serum stimulated cells with entrance into S phase retarded by five to six hours. If ouabain is added to serum stimulated cells at six hours, after the increase in K transport and K content have occurred, entrance into S phase is not entirely blocked. In cells stimulated with serum and 0.5 mM dBcAMP plus 1 mM theophylline simultaneously, entrance into S phase is greatly reduced as compared to serum stimulation only. However, the early and late changes in K flux and K content are not substantially altered. This indicates that the K transport events associated with G₁ and early S phase are not directly regulated by changes in cAMP levels which follow serum stimulation.     

Effects of External Potassium and Strophanthidin on Sodium Fluxes in Frog Striated Muscle

Unidirectional Na fluxes in isolated fibers from the frog''s semitendinosus muscle were measured in the presence of strophanthidin and increased external potassium ion concentrations. Strophanthidin at a concentration of 10^-5 M inhibited about 80 per cent of the resting Na efflux without having any detectable effect on the resting Na influx. From this it is concluded that the major portion of the resting Na efflux is caused by active transport processes. External potassium concentrations from 2.5 to 7.5 mM had little effect on resting Na efflux. Above 7.5 mM and up to 15 mM external K, the Na efflux was markedly stimulated; with 15 mM K the Na influx was 250 to 300 per cent greater than normal. On the other hand, Na influx was unchanged with 15 mM K. The stimulated Na efflux with the higher concentrations was not appreciably reduced when choline or Li was substituted for external Na, but was completely inhibited by 10^-5 M strophanthidin. From these findings it is concluded that the active transport of Na is stimulated by the higher concentrations of K. It is postulated that this effect on the Na "pump" is produced as a result of the depolarization of the muscle membranes and is related to the increased metabolism and heat production found under conditions of high external K.     

The Potassium Relations of Lemna minor L.: I. POTASSIUM UPTAKE AND PLANT GROWTH

A simple expression has been derived to predict the rate ofnet K uptake into exponentially growing plants of Lemna minor.Net uptake predictions are in good quantitative agreement withmeasurements of ‘steady-state’ K influx, indicatingthat, in the ‘steady state’, K movements in theplant are essentially undirectional and that efflux is small.This close matching of inward K movement to the demands of theexpanding tissue is temporarily disturbed if plants are transferredto media of different K status. Uptake rates in the ‘step-up’are initially enhanced and then fall gradually towards a new‘steady-state’ rate. In contrast, the ‘step-down’causes an initial depression of uptake and then rates increasegradually towards the new ‘steady-state’ rate. Itis argued that these changes in uptake rates are associatedwith alterations in the cytoplasmic K content.     

Potassium: a vital nutrient mediating stress tolerance in plants

Nutrients have been known to affect stress conditions, in fact, nutrient deprivations are stress conditions for plants itself. Likewise, three important nutrients Nitrogen (N), Phosphorus (P) and Potassium (K) mediates major stress responses in plants. Here, involvement of K has been discussed briefly in plant stress response along with its impact on plant development. K has been regarded as immensely important nutrient in agriculture, hence, its deficiency triggers various signaling cascades, finally enabling plants to activate stress adaptation responses. So far, K⁺ has been reported to play pivotal role in various abiotic stresses such as drought, cold, water stresses etc. However, the exact mechanism and interplay of these different abiotic stress regulation by K⁺ is not completely explored and demand further functional investigations. The in-depth understanding of components involved in K⁺ sensing, transport, and homeostasis will enable plant biologist to engineer crop varieties tolerant to abiotic stresses and nutrient deficient soil in near future.

     

Potassium and phosphorus uptake by corn genotypes grown in the field as influenced by root characteristics

Summary Root parameters of three corn (Zea mays L.) genotypes influencing P and K uptake were investigated in solution culture and field experiments. The data for these parameters were used to simulate P and K uptake by plants grown in the field using the Claassen-Barber model⁵. Root characteristics for ion influx, maximum rate of influx,Imax; Michaelis-Menten constant,Km; and minimum concentration of solution below which no further net influx occurs,Cmin were determined in solution culture. These kinetic parameters varied 2 to 3 fold among genotypes. Variations among genotypes were different for K than for P.Three corn genotypes were grown in the field and harvested 47, 54 and 68 days after emergence. Yield and root surface per plant increased about 3 fold during this time. At 47 days, 2/3 of the total root surface was in the top soil whereas 3 weeks later, it was less than 50%. Genotypes differed in distribution of roots between the topsoil and subsoil as well as in root surface per unit of shoot.K uptake predicted by the Claassen-Barber model was 2 to 3 times the observed. The overprediction could be related to high root density (length of root per unit soil volume) which indicated that competition between roots occurred that was not considered in the simulation model. The predicted P uptake (y) was correlated (r=0.91) to observed uptake (x) byy=0.98+0.67x, indicating underprediction of P uptake. The presence of root hairs may have been the cause of the underprediction. The calculated contribution of the subsoil to the observed uptake was 10% for K and 1% in the case of P. It was concluded that the plant parameters used to simulate nutrient uptake were rated accurately when allowance was made for root competition and presence of root hairs.Journal Paper No. 7608. Purdue University, Agric. Exp. Station, West Lafayette, IN 47907. Contribution from the Department of Agronomy. This research was supported in part by the Tennessee Valley Authority and the Deutsche Forschungsgemeinschaft.     

Energy-linked Potassium Influx as Related to Cell Potential in Corn Roots

Cell potentials and K⁺ (⁸⁶Rb) influx were determined for corn roots over a wide range of external K⁺ activity (K°) under control, anoxic, and uncoupled conditions. The data were analyzed using Goldman theory for the contribution of passive influx to total influx. For anoxic and uncoupled roots the K⁺ influx shows the functional relationship with K° predicted with constant passive permeability, although K⁺ permeability in uncoupled roots is about twice that of anoxic roots. In control roots the equation fails to describe K⁺ influx at low K°, but does so at high K°, with a gradual transition over the region where the electrical potential becomes equal to the equilibrium potential for K⁺ (ψ = E_K). In the low K° range, where net K⁺ influx is energetically uphill, participation of an energy-linked K⁺ carrier is indicated. In the high K° range, K⁺ influx becomes passive down the electrical gradient established by the cell potential. Since the cell potential includes a substantial electrogenic component, anoxia or uncoupling reduces passive influx.     

Potassium uptake efficiency and dynamics in the rhizosphere of maize (Zea mays L.), wheat (Triticum aestivum L.), and sugar beet (Beta vulgaris L.) evaluated with a mechanistic model

Plant species differ in nutrient uptake efficiency. With a pot experiment, we evaluated potassium (K) uptake efficiency of maize (Zea mays L.), wheat (Triticum aestivum L.), and sugar beet (Beta vulgaris L.) grown on a low-K soil. Sugar beet and wheat maintained higher shoot K concentrations, indicating higher K uptake efficiency. Wheat acquired more K because of a greater root length to shoot dry weight ratio. Sugar beet accumulated more shoot K as a result of a 3- to 4-fold higher K influx as compared to wheat and maize, respectively. Nutrient uptake model NST 3.0 closely predicted K influx when 250 mg K kg^?1 were added to the soil, but under-predicted K influx under low K supply. Sensitivity analysis showed that increasing soil solution K concentration (C_Li) by a factor of 1.6–3.5 or buffer power (b) 10- to 50-fold resulted in 100% prediction of K influx. When both maximum influx (I_max) and b were increased by a factor of 2.5 in maize and wheat and 25 in sugar beet, the model could predict measured K influx 100%. In general, the parameter changes affected mostly calculated K influx of root hairs, demonstrating their possible important role in plant K efficiency.     

Changes from High Potassium (HK) to Low Potassium (LK) in Bovine Red Cells

Red cells of newborn calves contain 105–110 mmole K⁺ and 1–5 mmole Na⁺ per liter of cells. As the animals age the K⁺ content decreases to a value of 25–30 mmole/liter of cells after about 60 days. At approximately the same time, the sodium content reaches a value of 60–70 mmole/liter. The time required for half change (t_½) is 35–37 days for both Na⁺ and K⁺. The activity of (Na + K)-adenosine triphosphatase (ATPase) and the influx of K⁴² and Rb⁸⁶ into the red cells are high at birth and are reduced to 5 and 15% of their original values, respectively, in mature animals. t_½ for both is of the order of 30–35 days. The membrane Mg-ATPase activity is also high at birth and is reduced with a t_½ of 28–32 days to a final value of about 20% of its activity at birth. Separation of red cells according to their age showed that, in animals at the age of transition, newly formed red cells contain a higher K/Na ratio and a higher active transport capacity than older red cells of the same animal. It is suggested that the changes observed are a reflection of the average age of the red cell population as the animal grows.     

Sodium Extrusion and Potassium Uptake in Guinea Pig Kidney Cortex Slices

Slices from the cortex corticis of the guinea pig kidney were immersed in a chilled solution without K and then reimmersed in warmer solutions. The Na and K concentrations and the membrane potential V_m were then studied as a function of the Na and K concentrations of the reimmersion fluid. It was found that Na is extruded from the cells against a large electrochemical potential gradient. Q₁₀ for net Na outflux was ∼2.5. At bath K concentrations larger than 8 mM the behavior of K was largely passive. At the outset of reimmersion (V_m > E_K) K influx seemed secondary to Na extrusion. Na extrusion would promote K entrance, being limited and requiring the presence of K in the bathing fluid. At bath K concentrations below 8 mM, K influx was up an electrochemical potential gradient. Thus a parallel active K uptake is apparent. Q₁₀ for net K influx was ∼2.0. Dinitrophenol inhibited net Na outflux and net K influx, Q₁₀ became <1.1 for both fluxes. The ratio between these fluxes varied. Thus at the outset of reimmersion the net Na outflux to net K influx ratio was >1. After 8 minutes it was <1.     

Equilibrium and Ion Exchange Characteristics of Potassium and Sodium Accumulation by Barley Roots

Potassium ion and Na⁺ influx and efflux rates into and from excised barley roots are compared with the maximum capacity of accumulation. Potassium ion and Na⁺ influx and efflux involve a cation exchange that is independent of simultaneous exchange of the accompanying anion. These exchange fluxes depend on the concentration and cation composition of the solutions from which they originate. Selective differences between K⁺ and Na⁺ fluxes are sufficient to account for a cationic distribution within the roots that differs markedly from that of the external solution and that persists for extended time periods. The accumulation maximum is a cation exchange equilibrium with the cation influx and efflux rates approaching equality. The equilibrium level is independent of the individual cation fluxes and the external solution concentration. It is a finite quantity which appears to be determined by the internal anion concentration including accumulated as well as endogenous anions.     

A Model for the Regulation of K Influx, and Tissue Potassium Concentrations by Negative Feedback Effects upon Plasmalemma Influx

By means of a modified Michaelis-Menten equation for K⁺ influx, which includes terms for root and external K⁺ concentrations (root [K⁺] and [K⁺]₀, respectively) it is possible to predict the manner in which short-term (perturbation) fluxes of K⁺ into roots of barley plants (Hordeum vulgare cv Fergus) vary with root [K⁺] and [K⁺]₀. Influx values derived from this equation were used to predict changes of root and shoot [K⁺] and K⁺ absorption rates (as functions of time and [K⁺]₀) from a knowledge of K⁺ efflux, relative growth rates of roots and shoots, and the partitioning of absorbed K⁺ between these organs. A microcomputer program was employed to model these changes in low-salt plants following transfer to solutions in which [K⁺]₀ was maintained at values ranging from 5 to 1000 millimoles per cubic meter. The model was operated on the basis of 10 minute absorption periods which provided data for continuous `updating' of tissue [K⁺]. The simulations were undertaken for periods corresponding to 30 days. During this time the model accurately predicted the manner in which K⁺ influx and root and shoot [K⁺] gradually approach values which are essentially independent of [K⁺]₀. The computer program was also used to predict the outcome of changing various external and internal parameters of the proposed regulatory system. The results of these simulations are discussed in the context of current models for negative feedback control of ion fluxes.     

Potassium Transport in Corn Roots : IV. Characterization of the Linear Component

A detailed examination was conducted on the linear, or first-order kinetic component for K⁺(⁸⁶Rb⁺) influx into root segments of both low- and high-salt grown corn seedlings (Zea mays [A632 × Oh 43]). In tissue from both low- and high-salt grown roots, replacement of Cl⁻ in the uptake solution by either SO₄²⁻, H₂PO₄⁻, or NO₃⁻ caused a significant (50-60%) and specific inhibition of the linear component of K⁺ influx. The anion transport inhibitor, 4,4′-diisothiocyano-2,2′-disulfonic acid, was found to abolish saturable Cl⁻ influx in corn roots while causing a significant (50-60%) and specific inhibition of the linear K⁺ uptake system; this inhibition was identical to that observed when Cl⁻ was replaced by other anions in the K⁺ uptake solution. Additionally, the quaternary ammonium cation, tetraethylammonium, which has been shown to block K⁺ channels in nerve axons, also caused a dramatic (70%) and specific inhibition of the linear component of K⁺ influx, but this was obtained only in high-salt roots. The reasons for this difference are discussed with respect to the differing abilities of low- and high-salt roots to absorb tetraethylammonium.

Our present results indicate that the linear component of K⁺ influx may occur by a passive process involving transmembrane K⁺ channels. Fluxes through these K⁺ channels may be partly coupled to a saturating Cl⁻ influx mechanism.

     

The effect of shoot-root ratio and temperature on K+ influx in rye

The temperature sensitivity of K⁺ influx into rye roots and root plasma membrane ATPase activity were compared in plants grown at different temperatures. It was shown that ATPase activity obeyed the Arrhenius relationship with temperature, whereas K⁺ influx into intact plants was linearly related to temperature and markedly influenced by shoot/root ratio. A model for acclimation of K⁺ influx to low temperatures based on the regulation of the K⁺ carrier mechanism by plant demand for K⁺ is described.