Active Transport Of Potassium Research Articles

What are the benefits of magnesium supplements?

Magnesium is one of the most abundant minerals in the body and is used in numerous cellular functions. It acts as a cofactor in enzymatic reactions, is an important part of energy production and glycolysis, contributes to bone development, and participates in the active transport of potassium and calcium ions across cell membranes.

Detection of Circulating Serum microRNA/Protein Complexes in ASD Using Functionalized Chips for an Atomic Force Microscope.

MicroRNAs, which circulate in blood, are characterized by high diagnostic value; in biomedical research, they can be considered as candidate markers of various diseases. Mature microRNAs of glial cells and neurons can cross the blood–brain barrier and can be detected in the serum of patients with autism spectrum disorders (ASD) as components of macrovesicles, macromolecular protein and low-density lipoprotein particles. In our present study, we have proposed an approach, in which microRNAs in protein complexes can be concentrated on the surface of AFM chips with oligonucleotide molecular probes, specific against the target microRNAs. MicroRNAs, associated with the development of ASD in children, were selected as targets. The chips with immobilized molecular probes were incubated in serum samples of ASD patients and healthy volunteers. By atomic force microscopy (AFM), objects on the AFM chip surface have been revealed after incubation in the serum samples. The height of these objects amounted to 10 nm and 6 nm in the case of samples of ASD patients and healthy volunteers, respectively. MALDI-TOF-MS analysis of protein components on the chip surface allowed us to identify several cell proteins. These proteins are involved in the binding of nucleic acids (GBG10, RT24, RALYL), in the organization of proteasomes and nucleosomes (PSA4, NP1L4), and participate in the functioning of the channel of active potassium transport (KCNE5, KCNV2).

Aldosterone up-regulates voltage-gated potassium currents and NKCC1 protein membrane fractions

Na+–K+–2Cl− Cotransporter (NKCC1) is a protein that aids in the active transport of sodium, potassium, and chloride ions across cell membranes. It has been shown that long-term systemic treatment with aldosterone (ALD) can enhance NKCC1 protein expression and activity in the aging cochlea resulting in improved hearing. In the present work, we used a cell line with confirmed NKCC1 expression to demonstrate that in vitro application of ALD increased outward voltage-gated potassium currents significantly, and simultaneously upregulated whole lysate and membrane portion NKCC1 protein expression. These ALD-induced changes were blocked by applying the mineralocorticoid receptor antagonist eplerenone. However, application of the NKCC1 inhibitor bumetanide or the potassium channel antagonist Tetraethyl ammonium had no effect. In addition, NKKC1 mRNA levels remained stable, indicating that ALD modulates NKCC1 protein expression via the activation of mineralocorticoid receptors and post-transcriptional modifications. Further, in vitro electrophysiology experiments, with ALD in the presence of NKCC1, K+ channel and mineralocorticoid receptor inhibitors, revealed interactions between NKCC1 and outward K+ channels, mediated by a mineralocorticoid receptor-ALD complex. These results provide evidence of the therapeutic potential of ALD for the prevention/treatment of inner ear disorders such as age-related hearing loss.

Role of active transport of potassium to leaves in the mechanisms of tolerance to salinity in common bean (Phaseolus vulgaris L.)

Biochemical markers are of great utility in screening for salt tolerance of crops. In common beans (Phaseolus vulgaris), lower levels of proline under stress have been associated with a better stress resistance of cultivars. In the present study, the responses to salinity have been analysed in six cultivars of common beans: four local landraces from Spain and two experimental lines from Cuba. Proline was used for ranking the relative tolerance of the cultivars, confirming a previous study which reported as more stress-tolerant two of the Spanish landraces. Total soluble sugars concentrations varied with treatments and between genotypes, but it was difficult to assess their role in stress tolerance of the analysed plants. Sodium concentration in leaves was the lowest in one of the two salt-resistant cultivars, and potassium did not vary or even increased under salt stress in all of them, except for the most susceptible one, where a drop of this cation was registered under 150 mM NaCl. Changes in malondialdehyde (MDA) contents did not indicate salt-induced membrane peroxidation resulting from secondary oxidative stress; consequently, accumulation of total phenolic compounds and flavonoids, as an antioxidant defence mechanism, was not detected. These results highlight the reliability of using proline as a biochemical marker of salt stress in common beans and the importance of the mechanism related to potassium transport to leaves in conferring stress tolerance to some common bean cultivars.

Model of fluid and solute shifts during hemodialysis with active transport of sodium and potassium.

BackgroundMathematical models are useful tools to predict fluid shifts between body compartments in patients undergoing hemodialysis (HD). The ability of a model to accurately describe the transport of water between cells and interstitium (Jv,ISIC), and the consequent changes in intracellular volume (ICV), is important for a complete assessment of fluid distribution and plasma refilling. In this study, we propose a model describing transport of fluid in the three main body compartments (intracellular, interstitial and vascular), complemented by transport mechanisms for proteins and small solutes.MethodsThe model was applied to data from 23 patients who underwent standard HD. The substances described in the baseline model were: water, proteins, Na, K, and urea. Small solutes were described with two-compartment kinetics between intracellular and extracellular compartments. Solute transport across the cell membrane took place via passive diffusion and, for Na and K, through the ATPase pump, characterized by the maximum transport rate, JpMAX. From the data we estimated JpMAX and two other parameters linked to transcapillary transport of fluid and protein: the capillary filtration coefficient Lp and its large pores fraction αLP. In an Expanded model one more generic solute was included to evaluate the impact of the number of substances appearing in the equation describing Jv,ISIC.ResultsIn the baseline model, median values (interquartile range) of estimated parameters were: Lp: 11.63 (7.9, 14.2) mL/min/mmHg, αLP: 0.056 (0.050, 0.058), and JpMAX: 5.52 (3.75, 7.54) mmol/min. These values were significantly different from those obtained by the Expanded model: Lp: 8.14 (6.29, 10.01) mL/min/mmHg, αLP: 0.046 (0.038, 0.052), and JpMAX: 16.7 (11.9, 25.2) mmol/min. The relative RMSE (root mean squared error)averaged between all simulated quantities compared to data was 3.9 (3.1, 5.6) %.ConclusionsThe model was able to accurately reproduce most of the changes observed in HD by tuning only three parameters. While the drop in ICV was overestimated by the model, the difference between simulations and data was less than the measurement error. The biggest change in the estimated parameters in the Expanded model was a marked increase of JpMAX indicating that this parameter is highly sensitive to the number of species modeled, and that the value of JpMAX should be interpreted only in relation to this factor.

The clock protein PERIOD is expressed in goblet cells of the larval midgut in the silkworm, Bombyx mori

Insects have both a circadian oscillatory system in peripheral tissues and a master clock in the central nervous system. However, the distribution of oscillator cells in the digestive system that is indispensable for nutrient intake has not been described in detail. This study immunohistochemically investigates cellular localization of the clock protein PERIOD in the larval midgut of the silkworm Bombyx mori (Lepidoptera: Bombycidae). PERIOD-like immunoreactivity was not uniformly distributed and was located exclusively in goblet cells of the midgut epithelium. The primary role of goblet cells is the active transport of potassium from hemolymph into the gut lumen. However, these cells might function as a peripheral clock that regulates the daily rhythms of physiological activities in the midgut of silkworm larvae.

Early recognition of reverse pseudohyperkalemia in heparin plasma samples during leukemic hyperleukocytosis can prevent iatrogenic hypokalemia

ObjectivesTo investigate the cause of apparent hyperkalemia in leukemic heparin plasma. Design and methodsLithium heparin plasma and serum samples from a patient with chronic lymphocytic leukemia (CLL) with hyperleukocytosis were transported by either a pneumatic tube system or manual transport and analyzed either immediately or after 4h. ResultsPneumatic tube transported samples resulted in higher plasma potassium levels than manually transported samples. Serum potassium was lower than plasma potassium, confirming the suspicion of “reverse” pseudohyperkalemia. Letting the pneumatic tube transported samples stand on the bench for 4h before centrifugation surprisingly resulted in decreased or unchanged plasma potassium. ConclusionsThe reverse pseudohyperkalemia in heparin plasma samples from a CLL patient was caused by pneumatic tube transport. Our results suggest extracellular leakage of potassium, followed by active transport of potassium into intact leukemic cells. This is the first Swedish case of reverse pseudohyperkalemia in a CLL patient, where clinical suspicion of false hyperkalemia and awareness of the phenomenon lead to a rapid laboratory diagnosis. The demonstration of reverse pseudohyperkalemia prevented potentially dangerous medical interventions, such as potassium lowering treatment.

Impact of improved potassium accumulation on pH homeostasis, membrane potential adjustment and survival of Corynebacterium glutamicum

Metal ion uptake is crucial for all living cells and an essential part of cellular bioenergetic homeostasis. In this study the uptake and the impact of the most abundant internal cation, potassium, were investigated in Actinobacteria, a group of high G + C Gram-positives with a number of prominent biotechnologically and medically important members. Genome analyses revealed a variety of different potassium uptake systems in this monophyletic group ranging from potassium channels common in virtually all Actinobacteria to different active carriers that were present predominantly in pathogenic members able to cope with various stress conditions. By applying Corynebacterium glutamicum as model system we provide experimental evidence that under optimal conditions a potassium channel is sufficient in bacteria for the maintenance of internal pH and membrane potential ensuring survival of cells under stress conditions. Under potassium limitation, however, viability of C. glutamicum was increased under acidic stress or during desiccation when a functional KtrAB potassium transporter from the pathogen Corynebacterium jeikeium was heterologously expressed. We provide experimental evidence that the KtrAB mediated enhanced potassium accumulation improved maintenance of internal pH and membrane potential. The results indicate that the occurrence of active potassium transport systems correlates with an improved potassium-dependent bioenergetic homeostasis and survival of bacterial cells under stress conditions.

Cell-specific signal transduction pathways regulating Na+-K+-ATPase. Focus on “Short-term effects of thyroid hormones on the Na+-K+-ATPase activity of chick embryo hepatocytes during development: focus on signal transduction”

the na+-k+-atpase is a complex integral membrane protein that carries out active transport of sodium and potassium across the cell plasma membrane, maintaining the ionic gradients. Each subunit has multiple isozymes that are expressed in tissue and developmental specific fashion. In addition to

Sulphonylurea sensitivity and enriched expression implicate inward rectifier K+ channels inDrosophila melanogasterrenal function

Insect Malpighian (renal) tubules are capable of transporting fluid at remarkable rates. Secondary active transport of potassium at the apical surface of the principal cell must be matched by a high-capacity basolateral potassium entry route. A recent microarray analysis of Drosophila tubule identified three extremely abundant and enriched K(+) channel genes encoding the three inward rectifier channels of Drosophila: ir, irk2 and irk3. Enriched expression of inward rectifier channels in tubule was verified by quantitative RT-PCR, and all three IRKs localised to principal cells of the main segment (and ir and irk3 to the lower tubule) by in situ hybridisation, suggesting roles both in primary secretion and reabsorption. A new splice form of irk2 was also identified. The role of inward rectifiers in fluid secretion was assessed with a panel of selective inhibitors of inward rectifier channels, the antidiabetic sulphonylureas. All completely inhibited fluid secretion, with IC(50)s of 0.78 mmol l(-1) for glibenclamide and approximately 5 mmol l(-1) for tolbutamide, 0.01 mmol l(-1) for minoxidil and 0.1 mmol l(-1) for diazoxide. This pharmacology is consistent with a lower-affinity class of inward rectifier channel that does not form an obligate multimer with the sulphonylurea receptor (SUR), although effects on non-IRK targets cannot be excluded. Glibenclamide inhibited fluid secretion similarly to basolateral K(+)-free saline. Radiolabelled glibenclamide is both potently transported and metabolised by tubule. Furthermore, glibenclamide is capable of blocking transport of the organic dye amaranth (azorubin S), at concentrations of glibenclamide much lower than required to impact on fluid secretion. Glibenclamide thus interacts with tubule in three separate ways; as a potent inhibitor of fluid secretion, as an inhibitor (possibly competitive) of an organic solute transporter and as a substrate for excretion and metabolism.

Neuronal function and alpha3 isoform of the Na/K-ATPase

The Na/K-ATPase is a complex of integral membrane proteins that carries out active transport of sodium and potassium across the cell plasma membrane, and maintains chemical gradients of these ions. The alpha subunit of the Na/K-ATPase has several isoforms that are expressed in a cell type- and tissue-dependent manner. In adult vertebrates, while kidney cells express mostly alpha1, muscle and glial cells -- alpha1 and alpha2, and sperm cells -- alpha1 and alpha4 isoforms of Na/K-ATPase, neurons may express alpha1, alpha2, alpha3 or any combination of these isoforms, and evidence suggests that neuronal type is the determining factor. The functional significance of multiple isoforms of the Na/K-ATPase and their non-uniform expression, and the link between neuron function and expression of a given isoform of the Na/K-ATPase in particular, remains unknown. Several hypotheses on this account were introduced, and in this work we will review the present status of these hypotheses, and their standing in application to recent data on the expression of isoforms of the Na/K-ATPase in the peripheral nervous system of vertebrate animals.

Active potassium transport across guinea-pig distal colon: action of secretagogues.

1. Adrenaline (5 microM) stimulated a K+ secretory current by 2.2 mu equiv h-1 cm-2 in isolated guinea-pig distal colonic epithelium. This secretory activity was inhibited entirely by addition of the loop diuretic bumetanide to the serosal solution. On-going K+ uptake via the absorptive pathway was unaltered by these changes. 2. Prostaglandin E2 (PGE2, 2 microM) stimulated electrogenic K+ secretion and Cl- secretion by 3.0 and 3.6 mu equiv h-1 cm-2, respectively. Serosal addition of bumetanide completely inhibited this K+ secretion but blocked only approximately 70% of Cl- secretion. The bumetanide-insensitive Cl- secretory current was dependent on the presence of Cl- and HCO3- in the bathing solutions. 3. Stimulation of electrogenic K+ secretion by PGE2 occurred with a half-maximal concentration of 4 nM, an affinity approximately 300 times higher than that for stimulation of Cl- secretion by PGE2. 4. Forskolin (10 microM) stimulated Cl- secretion by 4.9 mu equiv h-1 cm-2. The apparent K+ secretory rate was increased by only 1.5 mu equiv h-1 cm-2. A bumetanide-insensitive short-circuit current (ISC) was apparent and of the same size as that stimulated by PGE2. 5. Addition of the Ca2+ ionophore A23187 (10 microM), in the presence of indomethacin (1 microM) to reduce prostaglandin production, inhibited the K+ absorptive pathway by 40% and concurrently stimulated a small rate of electrogenic K+ secretion. 6. Active K+ absorption was inhibited by the addition of ouabain, omeprazole or SCH28080 to the mucosal solution. Both omeprazole and SCH28080 also stimulated a small negative ISC, consistent with electrogenic K+ secretion. 7. Association of K+ absorption, K+ secretion and Cl- secretion is indicated by similarities in transport mechanism and by secretagogue regulation. In particular, maximal rates of K+ secretory current require uptake via apical membrane K+ pumps. Such interrelations support a common cellular locus for these ion transport pathways.

Cation transport mediated by Na+,K(+)-adenosine triphosphatase in lymphoblastoma cells from patients with bipolar I disorder, their relatives, and unrelated control subjects.

In an investigation of cation transport in bipolar affective disorder, we have measured parameters related to Na+,K(+)-adenosine triphosphatase, the enzyme that carries out active transport of sodium and potassium, in lymphoblastoid cells cultured from patients with bipolar affective disorder, age-matched nonaffected family relatives, and unrelated control subjects. Patients had lower ion transport per cell and per transport enzyme site than did related or unrelated control subjects. The rate of transport per cell appeared higher in nonaffected relatives of patients than in unrelated control subjects, though this difference did not reach significance. These data suggest that abnormally regulated ion transport may be associated with bipolar affective disorder independently of clinical state.

Responsiveness to stress in major depression

In an investigation of cation transport in bipolar affective disorder, we have measmed parameters related to Na,K-ATPase, the enzyme that carties out active transport of sodium and potassium, in lymphoblastoid cells cultured from patients with bipolar affective disorder, age-matched nonaffected family relatives, and unrelated controls. Patients and their relafives were from well.characterized pedigrees of Old Order Amish. The use of iymphoblastoid cell lines largely eliminated possible effects of factom such as clinical state or recent drug treatment. Patients had lower ion mmsport per cell (271:1:40 pmol/10 e cells-min for patients vs 490 + 107 for related controls and 396 + 79 for unrelated controls; pc 0.001 by Kmskal-Wallis test) and per mmsport enzyme site (reduced by about 35% in patients, pc 0.02) than did related or unrelated controls. These data suggest that abnormally regulated ion transport may be associated with bipolar affective disorder independently of clinical state.

Effects of anoxia and inhibitors of energy metabolism on potassium and sodium homeostasis in neurons of gastropod mollusk

The effect of anoxia and inhibitors of energy metabolism on intracellular concentrations of potassium and sodium, membrane potentials, permeability, active and passive ouabain-sensitive transport of potassium (determined with86Rb) was studied in neurons of the freshwater planorbis mollusk (Planorbarius corneus). X-ray microanalysis showed that incubation of isolated ganglia in oxygen-free medium induced no change in intracellular concentrations of potassium and sodium. In the presence of cyanide, absorption of oxygen by the ganglia ceased, but accumulation of86Rb decreased insignificantly. The membrane potential and permeability did not depend on addition of cyanide. Desoxyglucose, an inhibitor of glycolysis, decreased86Rb accumulation more than cyanide did. In the presence of inhibitors of both glycolysis and respiration, which excluded the possibility of mutual compensation of oxidation and glycolytic sources of energy supply,86Rb accumulation decreased to the highest degree. A hypothesis was formulated on the paramount importance of glycolytic ATP for maintaining ion homeostasis of the nerve cells. The problem of functionally facilitated compartmentation of intracellular energy sources is discussed.

Modulation of active potassium transport by aldosterone

1. 1. The role of aldosterone on active potassium transport across lizard colon under voltage-clamped conditions has been investigated. 2. 2. Control colons exhibited no net potassium flux ( J k net ) despite of the existence of active opposite unidi ectional fluxes. 3. 3. An important net secretory potassium flux was found in short-circuited aldosterone-stimulated colons. 4. 4. Mucosal amiloride did not change ( J k net) either in control or aldosterone-stimulated colons. 5. 5. Luminal barium alters K + transport in a manner consistent with the presence of barium-sensitive conductances at the apical membrane of both control and aldosterone-treated colons. 6. 6. The effects of ouabain and barium on control and aldosterone-induced potassium flows were consistent with a model involving basolateral uptake by an Na +-K +-ATPase and conductive exit across the apical membrane. 7. 7. The stimulatory effect of aldosterone on potassium secretion is associated with parallel increases of both basolateral K + entry and the apical conductive pathway.

Regulation of active sodium and potassium transport in the distal colon of the rat. Role of the aldosterone and glucocorticoid receptors.

To determine whether mineralocorticosteroids and glucocorticosteroids have specific effects on colonic electrolyte transport, we compared the effect of aldosterone and RU 28362, a glucocorticoid receptor-specific agonist that does not bind to the aldosterone receptor, on unidirectional Na, Cl, and K fluxes across isolated mucosa of the rat distal colon. Continuous infusion of aldosterone for 7 d produced changes in four specific transport processes: induction of both active electrogenic, amiloride-sensitive sodium absorption and active electrogenic potassium secretion, enhancement of active electroneutral potassium absorption, and inhibition of electroneutral Na-Cl absorption, the predominant transport process in this epithelium. In contrast, continuous infusion of RU 28362 for 1-11 d produced a sustained increase in electroneutral Na-Cl absorption. This glucocorticoid receptor-specific agonist did not induce electrogenic sodium absorption nor affect either potassium absorption or secretion. These studies demonstrate that aldosterone (i.e., mineralocorticoid) and glucocorticoid receptors modulate separate and specific changes in active sodium and potassium transport. These results suggest that other glucocorticoids (e.g., dexamethasone, methylprednisolone) are not glucocorticoid receptor-specific and that their effects on electrogenic sodium absorption and potassium transport most likely represent the binding of these agonists to the aldosterone receptor.

86Rb(K) influx and [ 3H]ouabain binding by human platelets: Evidence for β-adrenergic stimulation of Na-K ATPase activity

Although active transport of potassium into human platelets has been demonstrated previously, there is hitherto no evidence that human platelets have an ouabain-inhibitable Na-K ATPase in their membrane. The present study demonstrates active rubidium (used as an index of potassium influx), 86Rb(K), influx into platelets, inhibitable by ouabain, and also demonstrates the presence of specific [ 3H]ouabain binding by the human platelet. This 86RB(K) influx was stimulated by adrenaline, isoprenaline, and salbutamol, but noradrenaline caused a mild inhibition. Active 86Rb(K) influx by platelets was inhibited markedly by timolol, mildly by atenolol, but not by phentolamine. Therefore, active 86Rb(K) influx in human platelets is enhanced by stimulation of β adrenoceptors of the β 2 subtype. The platelet may therefore replace the leukocyte in future studies of Na-K ATPase activity. This would be a considerable advantage in view of the ease and rapidity of preparation of platelets.

Potassium-proton symport in Neurospora: kinetic control by pH and membrane potential.

Active transport of potassium in K+-starved Neurospora was previously shown to resemble closely potassium uptake in yeast, Chlorella, and higher plants, for which K+ pumps or K+/H+-ATPases had been proposed. For Neurospora, however, potassium-proton cotransport was demonstrated to operate, with a coupling ratio of 1 H+ to 1 K+ taken inward so that K+, but not H+, moves against its electrochemical gradient (Rodriguez-Navarro et al., J. Gen. Physiol. 87:649-674). In the present experiments, the current-voltage (I-V) characteristic of K+-H+ cotransport in spherical cells of Neurospora has been studied with a voltage-clamp technique, using difference-current methods to dissect it from other ion-transport processes in the Neurospora plasma membrane. Addition of 5-200 microM K+ to the bathing medium causes 10-150 mV depolarization of the unclamped membrane, and yields a sigmoid I-V curve with a steep slope (maximal conductance of 10-30 microS/cm2) for voltages of -300 to -100 mV, i.e., in the normal physiologic range. Outside that range the apparent I-V curve of the K+-H+ symport saturates for both hyperpolarization and depolarization. It fails to cross the voltage axis at its predicted reversal potential, however, an effect which can be attributed to failure of the I-V difference method under reversing conditions. In the absence of voltage clamping, inhibitors-such as cyanide or vanadate-which block the primary proton pump in Neurospora also promptly inhibit K+ transport and K+-H+ currents. But when voltage clamping is used to offset the depolarizing effects of pump blockade, the inhibitors have no immediate effect on K+-H+ currents. Thus, the inhibition of K+ transport usually observed with these agents reflects the kinetic effect of membrane depolarization rather than any direct chemical action or the cotransport system itself. Detailed study of the effects of [K+]o and pHo on the I-V curve for K+-H+ symport has revealed that increasing membrane potential systematically decreases the apparent affinity of the transporter for K+, but increases affinity for protons (Km range: for [K+]o, 15-45 microM; for [H+]o, 10-35 nM). This behavior is consistent with two distinct reaction-kinetic models, in which (i) a neutral carrier binds K+ first and H+ last in the forward direction of transport, or (ii) a negatively charged carrier (-2) binds H+ first and K+ last.

K transport in upper portion of descending limbs of long-loop nephron from hamster.

By use of the in vitro microperfusion technique, we investigated potassium transport in the upper portion of the descending limb of the long-loop nephron isolated from hamster kidney. The net potassium flux determined by ultramicro-flame photometry was -0.63 +/- 1.84 pmol X mm-1 X min-1, a value that was not significantly different from zero. The salt permeability for KCl was calculated from the amount of potassium entering the tubular lumen when the concentration of potassium in the bath was increased by approximately 5 mM. The value was 38.9 +/- 1.9 X 10(-5) cm X s-1. The bidirectional fluxes of 86Rb were measured as indices of potassium fluxes. Flux coefficients from lumen-to-bath and bath-to-lumen were 51.2 +/- 9.2 and 48.8 +/- 13.5 X 10(-5) cm X s-1, respectively. These values were not significantly different, confirming that there was no net flux for 86Rb. In another series of five experiments, the lumen-to-bath 86Rb flux coefficients were 69.4 +/- 13.2 X 10(-5) cm X s-1 in the absence of unlabeled Rb, and 70.2 +/- 13.9 X 10(-5) cm X s-1 in the presence of 5 mM unlabeled Rb. The lumen-to-bath 42K flux coefficient measured in the same series of animals was 85.3 +/- 10.5 X 10(-5) cm X s-1 (n = 10), a value that is slightly higher than, but not significantly different from, that of 86Rb. These data show that active potassium transport may exist in this segment and that the passive permeability for potassium is very high.(ABSTRACT TRUNCATED AT 250 WORDS)