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 When a nerve is in a solution in which 75 per cent of the NaCl has been replaced by choline chloride or sucrose, the active K influx exceeds the active Na efflux, and the K content is maintained.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.     

Relationship between ion requirements for respiration and membrane transport in a marine bacterium.

Intact cells of the marine bacterium Alteromonas haloplanktis 214 oxidized NADH, added to the suspending medium, by a process which was stimulated by Na+ or Li+ but not K+. Toluene-treated cells oxidized NADH at three times the rate of untreated cells by a mechanism activated by Na+ but not by Li+ or K+. In the latter reaction, K+ spared the requirement for Na+. Intact cells of A. haloplanktis oxidized ethanol by a mechanism stimulated by either Na+ or Li+. The uptake of alpha-aminoisobutyric acid by intact cells of A. haloplanktis in the presence of either NADH or ethanol as an oxidizable substrate required Na+, and neither Li+ nor K+ could replace it. The results indicate that exogenous and endogenous NADH and ethanol are oxidized by A. haloplanktis by processes distinguishable from one another by their requirements for alkali metal ions and from the ion requirements for membrane transport. Intact cells of Vibrio natriegens and Photobacterium phosphoreum oxidized NADH, added externally, by an Na+-activated process, and intact cells of Vibrio fischeri oxidized NADH, added externally, by a K+-activated process. Toluene treatment caused the cells of all three organisms to oxidize NADH at much faster rates than untreated cells by mechanisms which were activated by Na+ and spared by K+.     

(Na,K)-pump: cellular role and regulation in nonexcitable cells

The (Na,K)-pump develops and maintains ionic gradients that are of fundamental importance for proper function of most animal cells. These gradients are utilized in the form of ionic leak pathways by a number of special and general cell processes (e.g., nerve conduction, nutrient transport, pH regulation). As the sodium gradient in particular energizes many vital cell processes, alterations in cell activity will often be manifest as changes in sodium entry. The (Na,K)-pump rate varies accordingly, in order to maintain balance between Na entry and exit thereby maintaining the potential energy of the cell. Acute changes in sodium influx are balanced by increases in activity of existing pump units, with only a small change in intracellular sodium concentration. This is possible because intracellular is normally poised on the steep limb of the concentration versus activity curve for the (Na,K)-pump, at a point well below maximal activity, allowing large increases in (Na,K)-pump rate with only small changes in sodium concentration. If the increase in sodium influx is prolonged, it appears that the cell responds by synthesizing new pumps, allowing intracellular sodium concentration to return to its original values. Though increases in (Na,K)-pump activity must be accompanied by increases in potassium leak rates, in the experiments we have presented, there does not appear to be direct functional coupling between (Na,K)-pump and the K leak pathways. In these situations the matching of active influx and passive efflux of K short-term appears to occur by mechanisms not directly related to (Na,K)-pump activation.     

Effect of alloxan diabetes on (Na+ + K+)-activated ATPase in brush border membrane of the mucosal cell of rat small intestine]

In order to elucidate a possible relationship between (Na+ + K+)-activated ATPase and intestinal absorption of actively transported monosaccharides enzyme activity was measured in mucosal cells from alloxan diabetic rats. The general effect of increasing capacity of active, Na+-dependent transport processes in diabetes mellitus is associated with a significantly enhanced (Na+ +K+)-activated ATPase activity in mucosal homogenate from diabetic animals. To study the localization of these effects within the cell we isolated purified brush borders and their substructures. To enable a comparison to be made between preparation procedures of diabetic and control animals the fractions were controlled by electronmicroscopy and by measuring the sucrase activity. In the purified brush border fraction of alloxan treated rats there was no significant increase in (Na+ + K+)-activated ATPase activity. Based on these results we conclude that the (Na+ + K+)-activated ATPase in the basolateral membranes was increased in alloxan diabetes, and it seems very likely that this enzyme is involved in the regulation of Na+-dependent transport processes.     

Comparative studies of volume restoration following cold-stress induced swelling in renal tissues--I. Effects of ouabain, K+ free medium, colchicine and cytochalasin B on rat and rabbit kidney cortex slices

1. Cold-stress-induced swelling in rabbit and rat kidney cortex slices cannot be due to the sole inhibition of a Na+/K+ exchange system. In these tissues indeed, ouabain induces no swelling and an exchange of Na+ for K+ or a l/l basis. Inhibition of K+ extrusion at low temperature has also to be taken into consideration. 2. Volume restoration at 27 degrees C after cold-stress-induced swelling is inhibited by ouabain in rabbit slices, not in rat ones. The inhibition in rabbit slices is concomitant with an increase in Na+ at levels higher than equilibrium with the external medium. 3. Volume restoration does not seem to implicate colchicine or cytochalasin B sensitive processes.     

Transport of sodium and potassium in intestinal epithelial cells

Transport properties of rabbit small intestinal mucosa were investigated $\text{in vitro}$ to characterize the process by which epithelial cells maintain normal Na and K gradients across the cell membrane. Active transport of Na from the cell proceeded at a faster rate in the presence of K; and active transport of K into the cell was stimulated by the presence of Na. Following preincubation at 0°C to reduce tissue K content, a greater transmural electrical potential difference (PD) and short-circuit current (I_sc) developed as the temperature was raised to 37°C if K was present in the bathing solution. The PD and I_sc, which generally reflect the rate of active Na transport in ileum under control conditions, increased immediately upon raising the K concentration in the serosal solution from 0 to 10 nM.The results present the first direct indication in mammalian intestine of an interdependence of the Na and K active transport processes which regulate the intracellular content of these cations.     

Influence of calcium on sodium and potassium absorption by fresh and aged bean stem slices

The influence of Ca on the aging processes of bean stem (Phaseolus vulgaris) slices and on the absorption of K and Na by fresh and aged slices was investigated. In the presence of Ca, fresh tissue showed a preferential Na uptake. The preference for Na over K resulted from a differential depressive effect of Ca on absorption of these two ions. In aged tissue Na uptake was also depressed, but K absorption was accelerated, with a net result of a much greater absorption of K than Na.     

Activation of the Na+/K+/Cl- cotransport system by reorganization of the actin filaments in Ehrlich ascites tumor cells.

Reorganization (disassembly) of the actin filaments in Ehrlich ascites tumor cells, either by hypotonic treatment in the presence of Ca2+ or by addition of cytochalasin B, results in activation of the Na+/K+/Cl- cotransport system. However, other regulatory processes, some of which may be dependent on an intact filament system, are responsible for the activation of the Na+/K+/Cl- cotransport system after cell shrinkage.     

Immunocytochemical localization of NA+, K+-ATPase in primary cultures of rat retina

Conditions have been established which allow growth of embryonic rat retinal cells in dissociated cell culture for up to one month. Na+, K+-ATPase localization was studied in both neuronal and mixed neuronal-glial (flat cell) cultures. High Na+, K+-ATPase-like-immunoreactivity was associated with plasma membranes of neuronal cell bodies and their processes. Markedly lower immunoreactivity was found in the underlying flat cells in mixed cultures. Staining was generally uniform over perikaryal plasma membranes and showed a bead-like appearance in neuronal processes, supporting previous studies in brain tissue which used histocytochemical procedures specific for the Na+, K+-ATPase. This system should be useful for examining distribution of the enzyme in developing nerve and glial cells and may help to resolve questions regarding Na+-K+ homeostasis by neurons and glia.     

Analysis of Spike Electrogenesis and Depolarizing K Inactivation in Electroplaques of Electrophorus electricus, L

Voltage clamp analyses, combined with pharmacological tools demonstrate the independence of reactive Na and K channels in electrically excitable membrane of eel electroplaques. Spike electrogenesis is due to Na activation and is eliminated by tetrodotoxin or mussel poison, or by substituting choline, K, Cs, or Rb for Na in the medium. The K channels remain reactive, but K activation is always absent, the electroplaques responding only with K inactivation. This is indicated by an increased resistance when the membrane is depolarized by more than about 30 mv. The resting resistance (1 to 5 ohm cm²) is dependent upon the ionic conditions, but when K inactivation occurs the resistance becomes about 10 ohm cm² in all conditions. K inactivation does not change the EMF significantly. The transition from low to high resistance may give rise to a negative-slope voltage current characteristic, and to regenerative inactivation responses under current clamp. The further demonstration that pharmacological K inactivation (by Cs or Rb) leaves Na activation and spike electrogenesis unaffected emphasizes the independence of the reactive processes and suggests different chemical compositions for the membrane structures through which they operate.     

Na+-dependent K+ uptake Ktr system from the cyanobacterium Synechocystis sp. PCC 6803 and its role in the early phases of cell adaptation to hyperosmotic shock

Transmembrane ion transport processes play a key role in the adaptation of cells to hyperosmotic conditions. Previous work has shown that the disruption of a ktrB/ntpJ-like putative Na(+)/K(+) transporter gene in the cyanobacterium Synechocystis sp. PCC 6803 confers increased Na(+) sensitivity, and inhibits HCO(3)(-) uptake. Here, we report on the mechanistic basis of this effect. Heterologous expression experiments in Escherichia coli show that three Synechocystis genes are required for K(+) transport activity. They encode an NAD(+)-binding peripheral membrane protein (ktrA; sll0493), an integral membrane protein, belonging to a superfamily of K(+) transporters (ktrB; formerly ntpJ; slr1509), and a novel type of ktr gene product, not previously found in Ktr systems (ktrE; slr1508). In E. coli, Synechocystis KtrABE-mediated K(+) uptake occurred with a moderately high affinity (K(m) of about 60 microm), and depended on both Na(+) and a high membrane potential, but not on ATP. KtrABE neither mediated Na(+) uptake nor Na(+) efflux. In Synechocystis sp. PCC 6803, KtrB-mediated K(+) uptake required Na(+) and was inhibited by protonophore. A Delta ktrB strain was sensitive to long term hyperosmotic stress elicited by either NaCl or sorbitol. Hyperosmotic shock led initially to loss of net K(+) from the cells. The Delta ktrB cells shocked with sorbitol failed to reaccumulate K(+) up to its original level. These data indicate that in strain PCC 6803 K(+) uptake via KtrABE plays a crucial role in the early phase of cell turgor regulation after hyperosmotic shock.     

Active K+ transport in Mycoplasma mycoides var. Capri. Net and unidirectional K+ movements.

Analysis of the cation composition of growing Mycoplasma mycoides var. Capri indicates that these organisms have a high intracellular K+ concentration (Ki: 200--300 mM) which greatly exceeds that of the growth medium, and a low Na+ concentration (Na+i: 20 mM). Unlike Na+i,K+i varies with cell aging. The K+ transport properties studied in washed organisms resuspended in buffered saline solution show that cells maintain a steady and large K+ concentration gradient across their membrane at the expense of metabolic energy mainly derived from glycolysis. In starved cells, K+i decreases and is partially compensated by a gain in Na+. This substitution completely reverses when metabolic substrate is added (K+ reaccumulation process). Kinetic analysis of K+ movement in cells with steady K+ level shows that most of K+ influx is mediated by an autologous K+-K+ exchange mechanism. On the other hand, during K+ reaccumulation by K+-depleted cells, a different mechanism (a K+ uptake mechanism) with higher transport capacity and affinity drives the net K+ influx. Both mechanisms are energy-dependent. Ouabain and anoxia have no effect on K+ transport mechanisms; in contrast, both processes are completely blocked by dicyclohexylcarbodiimide, an inhibitor of the Mg2+ -dependent ATPase activity.     

Resolution of the potassium ion pump in muscle fibers using barium ions

When frog sartorius muscles recover from Na enrichment in the presence of external K, net K entry into the fibers occurs by both passive movement and active inward transport via a K pump. Under normal conditions, it has not been possible to experimentally distinguish these processes. Fractionation into the flux components must be accomplished from inferences concerning the K conductance or permeability during a period of rapid Na extrusion. The best estimates indicate that 60-80% of the K entry occurs via the K pump. In the presence of Ba ions, the membrane permeability to K is very much reduced. Under these conditions, Na-enriched muscles underwent a normal recovery in the presence of external K, and the amount of inward K movement due to the K pump rose to over 90% of the total K entry. The characteristics of the K pump studied by this means were: (a) essentially complete inhition by 10(-4) M ouabain, (b) inhibition by [Na], (c) activation by [K] according to a rectangular hyperbola in the absence of [Na], (d) linear activation by [Na]i over a wide range in concentration, (e) zero or undetectably low pumping rate as [Na]i leads to O, (f) the number of Na ions actively transported per K ion actively transported is 1.4-1.7 normally and 1.1 in the presence of Ba.     

Mechanisms and consequences of Na+,K+-pump regulation by insulin and leptin.

Hormonal control of the Na+,K+-pump modulates membrane potential in mammalian cells, which in turn drives ion coupled transport processes and maintains cell volume and osmotic balance. Na+,K+-pump regulation is particularly important in the musculoskeletal, cardiovascular and renal systems. Decreased Na+,K+-pump activity can result in a rise in intracellular Na+ concentrations which in turn increase Na+/Ca2+ exchange, thereby raising intracellular Ca2+ levels. In cardiac and skeletal muscle, this could interfere with normal contractile activity. Similarly, in vascular smooth muscle the result would be resistance to vasodilation. Inhibition of the Na+,K+-pump can also reduce the driving force for renal tubular Na+ reabsorption, elevating Na+ excretion. By virtue of decreasing the membrane potential, thus allowing more efficient depolarization of nerve endings and by increasing intracellular Ca2+, inhibition of the Na+,K+-pump can increase nervous tone. The ability of insulin to stimulate the Na+,K+-pump in various cells and tissues, and the physiological significance thereof, have been well documented. Much less is known about the effect of leptin on the Na+,K+-pump. We have shown that leptin inhibits Na+,K+-pump function in 3T3-L1 fibroblasts. Defects in insulin and leptin action are associated with diabetes and obesity, respectively, both of which are commonly associated with cardiovascular complications. In this review we discuss the mechanisms of Na+,K+-pump regulation by insulin and leptin and highlight how, when they fail, they may contribute to the pathophysiology of hypertension associated with diabetes and obesity.     

Contribution of an electrogenic pump to the resting membrane polarization in a crustacean heart.

1. In the neurogenic heart of the isopod crustacean Porcellio dilatatus, external K+ removal depolarized the membrane (K0 effect) whereas subsequent restoration of K+ resulted in a rapid hyperpolarization (K1 effect). 2. The amplitude of the K1 effect depended on the duration of the prior K+ deprivation and on the subsequent K+ concentration. 3. The membrane resistance slightly increased during the K0 effect; during the K1 effect, it only returned to its control value. 4. Ouabain, cooling and replacement of external Na+ by Li+ also produced depolarization. 5. The K1 effect was suppressed by ouabain and markedly depressed by lowering the temperature to 4-6 degrees C. It was abolished if Li+ replaced Na+ during the prior privation of K+; moreover Li+ was unable to act as a substitute for external K+ in generating the K1 effect if used at equivalent concentration, but enhanced the effect at high concentration. 6. The findings are consistent with the presence of an electrogenic sodium pump in the myocardium of Porcellio contributing to the resting membrane potential. 7. Changes in the spontaneous rhythm observed during K0 and K1 are further suggestive of the presence of an electrogenic Na+ pump in the pacemaker neurons of the cardiac ganglion. Another explanation is also proposed. 8. The magnitude of the spontaneous contractions of the heart was increased during the K0 effect and markedly decreased during the K1 effect. An indirect effect of the changes in internal Na+ concentration on the contractile processes is suggested.     

Physiological screening for target site insensitivity and localization of Na(+)/K(+)-ATPase in cardenolide-adapted Lepidoptera

Cardenolides are toxic plant compounds which specifically inhibit Na(+)/K(+)-ATPase, an animal enzyme which is essential for many physiological processes, such as the generation of action potentials. Several adapted insects feeding on cardenolide-containing plants sequester these toxins for their own defence. Some of these insects were shown to possess Na(+)/K(+)-ATPases with a reduced sensitivity towards cardenolides (target site insensitivity). In the present study we screened five species of arctiid moths feeding on cardenolide-containing plants for target site insensitivity towards cardenolides using an in vitro enzyme assay. The derived dose response curves of the respective Na(+)/K(+)-ATPases were compared to the insensitive Na(+)/K(+)-ATPase of the monarch butterfly (Danaus plexippus). Na(+)/K(+)-ATPases of all arctiid species tested were highly sensitive to ouabain, a water-soluble cardenolide which is most widely used in laboratory studies. Nevertheless, we detected substantial amounts of cardenolides in the haemolymph of two of the arctiid species. In caterpillars of the sequestering arctiid Empyreuma pugione and of D. plexippus we localized Na(+)/K(+)-ATPase by immunohistochemistry and western blot (in D. plexippus). Both techniques revealed strong expression of the enzyme in the nervous tissue and indicated weak expression or even absence in other tissues tested. We conclude that instead of target site insensitivity the investigated arctiid species use a different strategy to tolerate cardenolides. Most plausibly, the perineurium surrounding the nervous tissue functions as a barrier which prevents cardenolides from reaching Na(+)/K(+)-ATPase in the ventral nerve cord.     

Quantification of ATP-producing and consuming processes of Ehrlich ascites tumour cells

ATP production of Ehrlich ascites tumour cells was estimated on the basis of their coupled respiration and lactate formation. ATP-consuming processes were assessed from the effects of selective inhibitors of RNA synthesis, protein synthesis and proteolysis, Na+/K+-ATPase on respiration. The extent of protein synthesis and proteolysis were also determined directly. From these values and those of the inhibition of respiration by selective inhibitors, a P/O ratio of 2.2 was calculated. About 75% of the total ATP consumption could be assigned to specific processes. The major ATP-consuming processes of tumour cells in an amino-acid-enriched medium, in which they are in an approximate steady state, are protein synthesis with about 30% of total ATP consumption, and Na+/K+-ATPase with about 20%, while RNA synthesis, ATP-dependent proteolysis and Ca2+-ATPase contribute about 10% each. In an amino-acid-free glucose medium, protein synthesis is reduced to a third, with a corresponding decrease of respiration, whereas the rate of the other ATP-consuming processes is unchanged.     

Cation Transport in Escherichia coli : I. Intracellular Na and K concentrations and net cation movement

Methods have been developed to study the intracellular Na and K concentrations in E. coli, strain K-12. These intracellular cation concentrations have been shown to be functions of the extracellular cation concentrations and the age of the bacterial culture. During the early logarithmic phase of growth, the intracellular K concentration greatly exceeds that of the external medium, whereas the intracellular Na concentration is lower than that of the growth medium. As the age of the culture increases, the intracellular K concentration falls and the intracellular Na concentration rises, changes which are related to the fall in the pH of the medium and to the accumulation of the products of bacterial metabolism. When stationary phase cells, which are rich in Na and poor in K, are resuspended in fresh growth medium, there is a rapid reaccumulation of K and extrusion of Na. These processes represent oppositely directed net ion movements against concentration gradients, and have been shown to be dependent upon the presence of an intact metabolic energy supply.     

Lactoferrin stimulates erythrocyte Na+/K+ -adenosine triphosphatase: effect of some modulators of membrane phosphorylation

We studied the effect of some modulators of signal transduction on the erythrocyte Na+/ K+-ATPase. Go6976 and Go6983 (protein kinase C inhibitors) showed a stimulatory effect and calyculin A (protein phosphatase inhibitor) exerted an inhibitory effect on the Na pump activity. Some of the tested modulators of cell-signaling [protein phosphatase(s), phosphodiesterase, calmodulin and some protein kinases] interfered with the lactoferrin (Lf) stimulatory effect on the sodium pump. Lf itself was able to modulate the effect of some agents upon the pump activity. Moreover, an additive effect of stimulation was found when Lf and some agents were used simultaneously. The summarized results showed that: (i) Lf upregulates the Na+/K+-ATPase in erythrocytes and facilitates the K+ influx into the erythrocytes; (ii) the effect of pump stimulation is mediated by phosphorylation processes. These results suggest a potential opportunity for using Lf alone or together with other agents as a stimulator of the erythrocyte Na+/K+-ATPase.     

四川红杉-岷江冷杉原始林凋落物K和Na的归还动态

以凋落物为载体的养分归还是生态系统物质循环的主要过程,也是林木生长所需养分的重要来源。相对于其他元素,钾(K)和钠(Na)在调节渗透压方面有着极其重要的作用,且淋溶作用强烈,极易流失,但缺乏必要关注。因此,以海拔3589 m高山峡谷区典型的四川红杉(Larix mastersiana)-岷江冷杉(Abies faxoniana)原始林为研究对象,在2015年9月到2016年8月期间调查了不同类型凋落物中K和Na元素在不同时期的归还特征。结果表明:1)凋落物K和Na元素年归还量分别为25.34和4.44 kg·hm^-2,表现为:凋落叶>凋落枝>其他凋落物>凋落皮>附生植物>繁殖器官。其中,凋落叶和枝占总凋落物K和Na元素归还的85%以上; 2)总凋落物、凋落叶和枝的K、Na元素归还动态呈现"双峰型",最大值出现在5月和10月;凋落皮和繁殖器官K、Na元素归还动态呈现"单峰型",最大值出现在5月; 3)乔木层树种凋落物的K和Na元素呈现"双峰型"的归还模式,最大值在5月和10月,灌木树种凋落物的K和Na元素呈现"单峰型"归还模式,最大值在...