Effects of Inhibitors of Active Transport on 22Na and 42K Movements and on Nucleotide Levels in Rat Uteri at 25 °C

1971 ◽  
Vol 49 (3) ◽  
pp. 178-204 ◽  
Author(s):  
E. E. Daniel ◽  
Kathleen Robinson
Keyword(s):  
Water Content ◽  
Sodium Pump ◽  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Compartment Model ◽  
Sodium Influx ◽  
Na Pump ◽  
Cellular Fraction ◽  
Na Efflux ◽  
External K

The effects of inhibitors of the sodium pump (K-free solution or ouabain) on Na and K fluxes, water content, and adenine nucleotides were studied in rat uterine horns. The same four-compartment model [Formula: see text] was applied to K efflux as to Na efflux. However, for K efflux the larger cellular fraction, C, contained 70% of tissue K, and D could usually be ignored. K-free solutions did not alter ATP, ADP, or AMP levels, and had only small effects on efflux of Na from fresh or Na-rich tissues. In these tissues ouabain decreased Na efflux from the faster, larger cellular fraction (BK2 ↓) without affecting adenine nucleotide levels. K-free solutions or ouabain increased sodium influx, but neither inhibitor caused swelling. The inhibitors decreased water content of fresh tissues and failed to interfere with extrusion of water accompanying Na from Na-rich tissues. In addition to the ouabain-sensitive pump, another Na pump insensitive to K levels or ouabain was postulated to be responsible for control of cell volume and part of Na efflux in uteri. The K efflux was increased by ouabain, but not by K-free solutions. The K influx was decreased by K-free solutions, but was not affected by ouabain. Increasing external K (concentrations of 18.4 mM and higher) did not diminish the effects of ouabain on Na or K efflux. Neither cocaine nor adrenaline prevented the effect of ouabain on the Na or K efflux or on the increased Na influx. No evidence was obtained that a Na exchange–diffusion system operated in K-free solutions. It was concluded that the accepted views about the operation of a Na pump involving (Na+ + K+)-activated, ouabain-sensitive ATPase have to be modified for rat uteri.

Effects of Inhibitors of Metabolism on Adenine Nucleotides and on 22Na and 42K and Net Movements in Rat Uteri at 25 °C

1971 ◽  
Vol 49 (3) ◽  
pp. 205-239 ◽  
Author(s):  
E. E. Daniel ◽  
Kathleen Robinson
Keyword(s):  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Atp Depletion ◽  
Atp Content ◽  
Sodium Efflux ◽  
Cellular Fraction ◽  
Glucose Depletion ◽  
Na Efflux ◽  
Per Se

The effects of 10−3 M iodoacetate (IAA) and (or) 10−3 M dinitrophenol (DNP) on Na and K fluxes and contents and on adenine nucleotide levels of isolated rat uterine horns were studied. Early 22Na efflux was slightly increased by DNP in the fresh and Na-rich tissues. IAA and DNP alone or together reduced 22Na efflux from the larger cellular fraction (No. 2) in both fresh and Na-rich tissues. 22Na efflux from the smaller cellular fraction (No. 3) was accelerated by IAA and by DNP in Na-rich tissues. DNP increased 22Na influx in both types of tissue and caused net Na gain and K loss. In fresh tissues IAA or IAA plus DNP accelerated 22Na influx, but slowed this influx in Na-rich tissues. In fresh tissues the ATP content was reduced by 50% by DNP. After a 60-min exposure with IAA and a 15- to 20-min exposure with IAA plus DNP, the ATP levels were negligible. The onsets of action of IAA or of IAA plus DNP on Na fluxes were correlated with ATP depletion, but early acceleration of 22Na efflux by DNP was not. In fresh tissues 42K influx was slightly decreased at the time of ATP depletion and the influx was further slowed as tissue potassium was replaced by sodium. IAA plus DNP increased K efflux in 10 min and IAA alone increased K efflux after 100 min. Thus K flux changes were not well correlated with ATP depletion. Substitution of K for all the sodium in the bathing media did not alter the quality of the effects of IAA or IAA plus DNP on sodium efflux. When prolonged glucose depletion eliminated ATP and ADP, the effects of IAA could not be duplicated. But IAA alone, or with DNP, still caused alterations in the 22Na efflux. Therefore IAA acted on ion fluxes by a mechanism other than ATP depletion. Both fresh and Na-rich tissues swelled after ATP depletion. An effect on internal osmotic pressure rather than ATP-depletion per se was postulated. Other studies showed that Na-rich tissues were resistant to shrinking by hypertonic sucrose and became more so secondarily after ATP depletion because of increased sucrose permeability. Evidence from studies of swelling, as well as flux data, suggested that at least two Na pumps were present. Both were ATP-dependent. One was ouabain-sensitive and exchanged Na for K, while the other was ouabain-insensitive and controlled movement of Na with water.

scholarly journals Strophanthidin-sensitive sodium fluxes in metabolically poisoned frog skeletal muscle.

1976 ◽  
Vol 68 (4) ◽  
pp. 405-420 ◽  
Author(s):  
B G Kennedy ◽  
P De Weer
Keyword(s):  
Skeletal Muscle ◽  
Membrane Potential ◽  
Human Erythrocyte ◽  
Sodium Pump ◽  
Giant Axon ◽  
Squid Giant Axon ◽  
Frog Skeletal Muscle ◽  
Na Efflux ◽  
Unidirectional Fluxes ◽  
External K

Strophanthidin-sensitive and insensitive unidirectional fluxes of Na were measured in fog sartorius muscles whose internal Na levels were elevated by overnight storage in the cold. ATP levels were lowered, and ADP levels raised, by metabolic poisoning with either 2,4-dinitrofluorobenzene or iodoacetamide. Strophanthidin-sensitive Na efflux and influx both increased after poisoning, while strophanthidin-insensitives fluxes did not. The increase in efflux did not require the presence of external K but was greatly attenuated when Li replaced Na as the major external cation. Membrane potential was not markedly altered by 2,4-dinitrofluorobenzene. These observations indicate that the sodium pump of frog skeletal muscle resembles that of squid giant axon and human erythrocyte in its ability to catalyze Na-Na exchange to an extent determined by intracellular ATP/ADP levels.

scholarly journals The influence of external sodium ions on the sodium pump in erythrocytes

1968 ◽  
Vol 109 (3) ◽  
pp. 369-374 ◽  
Author(s):  
R. N. Priestland ◽  
R. Whittam
Keyword(s):  
Adenosine Triphosphatase ◽  
Sodium Pump ◽  
Erythrocyte Membranes ◽  
Sodium Ions ◽  
Adenosine Triphosphatase Activity ◽  
Sensitive Site ◽  
Na Efflux ◽  
Ion Movements ◽  
K Concentration ◽  
External K

1. A study has been made of the interaction between Na+ and K+ on the adenosine triphosphatase activity of erythrocyte ‘ghosts’, and on the K+ influx and Na+ efflux of intact erythrocytes. The adenosine triphosphatase activity and the ion movements were greater at a low external K+ concentration in the absence of Na+ than they were in the presence of 150mm-Na+. The inhibition by external Na+ of K+ influx had an inhibitory constant of 5–10mm. 2. Activation by K+ of kidney microsomal adenosine triphosphatase was retarded by Na+, and activation by Na+ was retarded by K+. Fragmented erythrocyte membranes behaved similarly. 3. These observations suggest that there is competition between Na+ and K+ at the K+-sensitive site of the membrane.

scholarly journals Sodium and potassium fluxes and membrane potential of human neutrophils: evidence for an electrogenic sodium pump.

1982 ◽  
Vol 79 (3) ◽  
pp. 453-479 ◽  
Author(s):  
L Simchowitz ◽  
I Spilberg ◽  
P De Weer
Keyword(s):  
Membrane Potential ◽  
Sodium Pump ◽  
Membrane Conductance ◽  
Human Neutrophils ◽  
Constant Field ◽  
Electrogenic Sodium Pump ◽  
Na Efflux ◽  
The Individual ◽  
Sodium And Potassium ◽  
External K

Sodium and potassium ion contents and fluxes of isolated resting human peripheral polymorphonuclear leukocytes were measured. In cells kept at 37 degrees C, [Na]i was 25 mM and [K]i was 120 mM; both ions were completely exchangeable with extracellular isotopes. One-way Na and K fluxes, measured with 22Na and 42K, were all approximately 0.9 meq/liter cell water . min. Ouabain had no effect on Na influx or K efflux, but inhibited 95 +/- 7% of Na efflux and 63% of K influx. Cells kept at 0 degree C gained sodium in exchange for potassium ([Na]i nearly tripled in 3 h); upon rewarming, ouabain-sensitive K influx into such cells was strongly enhanced. External K stimulated Na efflux (Km approximately 1.5 mM in 140-mM Na medium). The PNa/PK permeability ratio, estimated from ouabain insensitive fluxes, was 0.10. Valinomycin (1 microM) approximately doubled PK. Membrane potential (Vm) was estimated using the potentiometric indicator diS-C3(5); calibration was based on the assumption of constant-field behavior. External K, but not Cl, affected Vm. Ouabain caused a depolarization whose magnitude dependent on [Na]i. Sodium-depleted cells became hyperpolarized when exposed to the neutral exchange carrier monensin; this hyperpolarization was abolished by ouabain. We conclude that the sodium pump of human peripheral neutrophils is electrogenic, and that the size of the pump-induced hyperpolarization is consistent with the membrane conductance (3.7-4.0 microseconds/cm2) computed from the individual K and Na conductances.

The Platelet of the Newborn Infant – Adenine Nucleotide Metabolism and Release

1980 ◽  
Vol 43 (02) ◽  
pp. 099-103 ◽  
Author(s):  
J M Whaun ◽  
P Lievaart ◽  
Keyword(s):  
Newborn Infant ◽  
Adenine Nucleotide ◽  
Platelet Rich Plasma ◽  
Adenine Nucleotides ◽  
Newborn Infants ◽  
Specific Radioactivity ◽  
Nucleotide Metabolism ◽  
Breakdown Product ◽  
Term Infants ◽  
Adenine Nucleotide Metabolism

SummaryBlood from normal full term infants, mothers and normal adults was collected in citrate. Citrated platelet-rich plasma was prelabelled with 3H-adenine and reacted with release inducers, collagen and adrenaline. Adenine nucleotide metabolism, total adenine nucleotide levels and changes in sizes of these pools were determined in platelets from these three groups of subjects.At rest, the platelet of the newborn infant, compared to that of the mother and normal adult, possessed similar amounts of adenosine triphosphate (ATP), 4.6 ± 0.2 (SD), 5.0 ± 1.1, 4.9 ± 0.6 µmoles ATP/1011 platelets respectively, and adenosine diphosphate (ADP), 2.4 ± 0.7, 2.8 ± 0.6, 3.0 ± 0.3 umoles ADP/1011 platelets respectively. However the marked elevation of specific radioactivity of ADP and ATP in these resting platelets indicated the platelet of the neonate has decreased adenine nucleotide stores.In addition to these decreased stores of adenine nucleotides, infant platelets showed significantly impaired release of ADP and ATP on exposure to collagen. The release of ADP in infants, mothers, and other adults was 0.9 ± 0.5 (SD), 1.5 ± 0.5, 1.5 ± 0.1 umoles/1011 platelets respectively; that of ATP was 0.6 ± 0.3, 1.0 ± 0.1,1.3 ± 0.2 µmoles/1011 platelets respectively. With collagen-induced release, platelets of newborn infants compared to those of other subjects showed only slight increased specific radioactivities of adenine nucleotides over basal levels. The content of metabolic hypoxanthine, a breakdown product of adenine nucleotides, increased in both platelets and plasma in all subjects studied.In contrast, with adrenaline as release inducer, the platelets of the newborn infant showed no adenine nucleotide release, no change in total ATP and level of radioactive hypoxanthine, and minimal change in total ADP. The reason for this decreased adrenaline reactivity of infant platelets compared to reactivity of adult platelets is unknown.Infant platelets may have different membranes, with resulting differences in regulation of cellular processes, or alternatively, may be refractory to catecholamines because of elevated levels of circulating catecholamines in the newborn period.

The Effect of Temperature on Sodium Movements in Rat Uteri and a Model for Control of their Ion Content

1971 ◽  
Vol 49 (3) ◽  
pp. 240-262 ◽  
Author(s):  
E. E. Daniel ◽  
Kathleen Robinson
Keyword(s):  
Atp Depletion ◽  
Rate Coefficients ◽  
Effect Of Temperature ◽  
Ion Content ◽  
Substantial Fraction ◽  
Cellular Fraction ◽  
Diffusion Controlled ◽  
Na Efflux ◽  
Pinocytotic Vesicles ◽  
Na Uptake

The uptake and efflux of 22Na was studied in isolated rat uterine horns (both fresh and Na-rich) at 5, 15, 25, and 37 °C. Reduction of temperature from 37 °C to 25 or to 15 °C reduced 22Na uptake into, and efflux from, both the extracellular space and cells to the degree expected of a diffusion-controlled process (Q10 < 2). Reduction of the temperature to 5 °C during uptake into Na-rich horns revealed that a substantial fraction of cellular sodium became less exchangeable. At 5 °C, 22Na efflux was also markedly reduced, more than from ouabain or ATP depletion. Analysis of this change by curve-peeling and by reducing the temperature at various stages of efflux suggested that the main cause was a shift of 22Na from the larger, faster cellular fraction (No. 2) to the slower cellular fraction (No. 3). Bound 22Na was also markedly increased. The rate coefficients from curve-peeling for both cellular fractions were decreased. Radioactivity still in fraction 2 at 5 °C emerged at a rate of about half that at 15 °C. However, an overall coefficient for efflux of 22Na which would have emerged in fraction 2 at 15 or 25 °C showed that the Q10 for 22Na efflux between 5 and 15 °C was about 15. Tissues did not swell when they gained sodium at 5 °C. The effects of ouabain to increase 22Na influx and 42K efflux were eliminated at 5 °C. The effects of ATP depletion by iodoacetate and dinitrophenol to decrease 22Na efflux and to increase 22Na uptake, K loss, and swelling were reduced at 5 °C. Prior ATP depletion altered but did not prevent the marked reduction of efflux by cooling to 5 °C. Efflux of lithium, but not of potassium, was markedly slowed at 5 °C. K-free solutions still increased 22Na uptake at 5 °C. A model involving pinocytotic vesicles to explain these and earlier results was postulated.

scholarly journals L-Arginine Intake Effect on Adenine Nucleotide Metabolism in Rat Parenchymal and Reproductive Tissues

2012 ◽  
Vol 2012 ◽  
pp. 1-4 ◽  
Author(s):  
G. Kocic ◽  
J. Nikolic ◽  
T. Jevtovic-Stoimenov ◽  
D. Sokolovic ◽  
H. Kocic ◽  
...  
Keyword(s):  
Adenosine Deaminase ◽  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Tissue Growth ◽  
Nucleotide Metabolism ◽  
Amp Deaminase ◽  
Male Impotence ◽  
Adenine Nucleotide Metabolism ◽  
Adenosine Concentration ◽  
Male Wistar Rats

L-arginine is conditionally essetcial amino acid, required for normal cell growth, protein synthesis, ammonia detoxification, tissue growth and general performance, proposed in the treatment of men sterility and prevention of male impotence. The aim of the present paper was to estimate the activity of the enzymes of adenine nucleotide metabolism:5′-nucleotidase (5′-NU), adenosine deaminase (ADA), AMP deaminase, and xanthine oxidase (XO), during dietary intake of L-arginine for a period of four weeks of male Wistar rats. Adenosine concentration in tissues is maintained by the relative activities of the adenosine-producing enzyme,5′-NU and the adenosine-degrading enzyme-ADA adenosine deaminase. Dietary L-arginine intake directed adenine nucleotide metabolism in liver, kidney, and testis tissue toward the activation of adenosine production, by increased5′-NU activity and decreased ADA activity. Stimulation of adenosine accumulation could be of importance in mediating arginine antiatherosclerotic, vasoactive, immunomodulatory, and antioxidant effects. Assuming that the XO activity reflects the rate of purine catabolism in the cell, while the activity of AMP deaminase is of importance in ATP regeneration, reduced activity of XO, together with the increased AMP-deaminase activity, may suggest that adenine nucleotides are presumably directed to the ATP regenerating process during dietary L-arginine intake.

Characteristics of active Na transport in intact cardiac cells

1979 ◽  
Vol 236 (2) ◽  
pp. H189-H199 ◽  
Author(s):  
H. G. Glitsch
Keyword(s):  
Heart Muscle ◽  
Muscle Cells ◽  
Cardiac Cells ◽  
Na Transport ◽  
Concentration Gradients ◽  
Experimental Conditions ◽  
Na Pump ◽  
Na Efflux ◽  
Heart Muscle Cells ◽  
K Concentration

An active Na transport maintains the Na and K concentration gradients across the cell membrane of many cells and restores them following excitation. Heart muscle cells display frequent electrical discharges and thus the cardiac Na pump is of fundamental functional significance. Some methods for studying active Na transport are described. The active Na efflux from heart muscle cells is activated by an increase in the intracellular Na and the extracellular K concentration. The linkage between active Na efflux and active K influx varies widely according to the experimental conditions. The cardiac Na pump is electrogenic and can contribute directly to the membrane potential of the cells. The effects of active Na transport on contraction and intercellular coupling in myocardium are discussed.

scholarly journals Effect of adenine nucleotides on myo-inositol-1,4,5-trisphosphate-induced calcium release

1997 ◽  
Vol 325 (3) ◽  
pp. 661-666 ◽  
Author(s):  
Ludwig MISSIAEN ◽  
Jan B. PARYS ◽  
Humbert DE SMEDT ◽  
Ilse SIENAERT ◽  
Henk SIPMA ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Binding Site ◽  
Calcium Release ◽  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Release Process ◽  
A7r5 Cells ◽  
Channel Opening ◽  
Inositol 1,4,5 Trisphosphate

The effects of a whole series of adenine nucleotides on Ins(1,4,5)P3-induced Ca2+ release were characterized in permeabilized A7r5 smooth-muscle cells. Several adenine nucleotides activated the Ins(1,4,5)P3 receptor. It was observed that 3′-phosphoadenosine 5′-phosphosulphate, CoA, di(adenosine-5′)tetraphosphate (Ap4A) and di(adenosine-5′)pentaphosphate (Ap5A) were more effective than ATP. Ap4A and Ap5A also interacted with a lower EC50 than ATP. In order to find out how these adenine nucleotides affected Ins(1,4,5)P3-induced Ca2+ release, we have measured their effect on the response of permeabilized A7r5 cells to a progressively increasing Ins(1,4,5)P3 concentration. Stimulatory ATP and Ap5A concentrations had no effect on the threshold Ins(1,4,5)P3 concentration for initiating Ca2+ release, but they stimulated Ca2+ release in the presence of supra-threshold Ins(1,4,5)P3 concentrations by increasing the co-operativity of the release process. Inhibition of the Ins(1,4,5)P3-induced Ca2+ release at higher ATP concentrations was associated with a further increase in co-operativity and also with a shift in threshold towards higher Ins(1,4,5)P3 concentrations. ATP had no effect on the non-specific Ca2+ leak in the absence of Ins(1,4,5)P3. We conclude that the adenine-nucleotide-binding site can be activated by many different adenine nucleotides. Binding of these compounds to the transducing domain of the Ins(1,4,5)P3 receptor increases the efficiency of transmitting Ins(1,4,5)P3 binding to channel opening. The inhibition by high ATP concentrations is exerted at a different site, related to Ins(1,4,5)P3 binding.

scholarly journals The effect of butacaine on adenine nucleotide binding and translocation in rat liver mitochondria

1975 ◽  
Vol 148 (3) ◽  
pp. 527-531 ◽  
Author(s):  
D R Fayle ◽  
G J Barritt ◽  
F L Bygrave
Keyword(s):  
Rat Liver ◽  
Local Anaesthetic ◽  
Binding Sites ◽  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Nucleotide Binding ◽  
Local Anaesthetics ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Mitochondrial Inner Membrane

The effect of the local anaesthetic, butacaine, on adenine nucleotide binding and translocation in rat liver mitochondria partially depleted of their adenine nucleotide content was investigated. The range of butacaine concentrations that inhibit adenine nucleotide translocation and the extent of the inhibition are similar to the values obtained for native mitochondria. Butacaine does not alter either the total number of atractyloside-sensitive binding sites of depleted mitochondria, or the affinity of these sites for ADP or ATP under conditions where a partial inhibition of the rate of adenine nucleotide translocation is observed. The data are consistent with an effect of butacaine on the process by which adenine nucleotides are transported across the mitochondrial inner membrane rather than on the binding of adenine nucleotides to sites on the adenine nucleotide carrier. The results are briefly discussed in relation to the use of local anaesthetics in investigations of the mechanism of adenine nucleotide translocation.

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