The Energy Metabolism of Novikoff Ascites Hepatoma Cells. I. The Effects of Glucose on the Intracellular Level of Adenine Nucleotides

1971 ◽  
Vol 49 (6) ◽  
pp. 686-694 ◽  
Author(s):  
J. W. Gurd ◽  
P. G. Scholefield
Keyword(s):  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Rapid Decrease ◽  
Intracellular Level ◽  
Atp Content ◽  
Ascites Tumor ◽  
Ascites Hepatoma ◽  
Novikoff Hepatoma ◽  
Adenosine Phosphates ◽  
Layer Chromatography

When freshly prepared Novikoff hepatoma ascites tumor cells were incubated with glucose there was a rapid decrease in the ATP content of the cells. This was initially accompanied by an increased level of ADP but in the interval 6–10 min after the addition of glucose only 40% of the original adenine nucleotide content could be accounted for as adenosine phosphates. Analysis by thin-layer chromatography indicated breakdown via inosine 5′-phosphate (IMP), inosine, and hypoxanthine. Continued incubation led to resynthesis from hypoxanthine to IMP and thence to the adenine nucleotides.

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 The relation of sodium and potassium ion transport to the respiration and adenine nucleotide content of liver slices treated with inhibitors of respiration

1972 ◽  
Vol 129 (2) ◽  
pp. 427-438 ◽  
Author(s):  
G. D. V. Van Rossum
Keyword(s):  
Ion Transport ◽  
Concentration Ratio ◽  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Critical Value ◽  
Endogenous Respiration ◽  
Atp Content ◽  
Nucleotide Content ◽  
Liver Slices ◽  
Potassium Ion Transport

1. The dependence of the net transport of Na+and K+by rat liver on the respiration has been determined by incubating slices in the presence of varying concentrations of respiratory inhibitors. 2. Neither the rate of net transport nor the total amount of each ion transported was inhibited unless the rate of endogenous respiration was decreased below a critical value of about 330mmol of O2/h per kg of protein (i.e. 50% of the total endogenous respiration). 3. The uninhibited rate of respiration could be varied over a twofold range (380–770mmol of O2/h per kg of protein) by the use of different substrates, but the critical value for the onset of transport inhibition was quite constant (290–360mmol/h per kg of protein) under these different conditions. 4. Slices incubated at 38°C without inhibitors showed an increase of their ATP content and the concentration ratio ATP/ADP. The final ATP content and concentration ratio, ATP/ADP, of slices treated with different concentrations of inhibitors were closely related to the rate of respiration. 5. The increased ATP content of the control slices during incubation was equal to the increase of total adenine nucleotides. At increasing degrees of respiratory inhibition the relative contributions of ADP and AMP to the total adenine nucleotide content increased. 6. The critical rate of respiration for the onset of inhibition of ion transport and the corresponding contents of adenine nucleotides provide estimates of the maximal values of certain parameters of energy metabolism required for the support of alkali-cation transport in the liver slices.

Adenine Nucleotide Metabolism of Human Platelets

1971 ◽  
Vol 25 (02) ◽  
pp. 223-233
Author(s):  
M Murakami ◽  
K Odake
Keyword(s):  
Adenine Nucleotide ◽  
Platelet Rich Plasma ◽  
Adenine Nucleotides ◽  
Human Platelets ◽  
Nucleotide Metabolism ◽  
The Other ◽  
Supernatant Fraction ◽  
Two Dimensional ◽  
Adenine Nucleotide Metabolism ◽  
Layer Chromatography

SummaryAfter platelet-rich plasma was incubated with radioactive adenine, radioactive adenine nucleotides in platelets were separated by two-dimensional thin-layer chromatography.Radioactive adenine was selectively incorporated into adenine nucleotides. Gradual decomposition of labelled nucleotides was observed after longer period of incubation. Radioactive ATP, ADP, AMP, IMP, and hypoxanthine were separated from PCA extract of platelets. On the other hand, radioactive adenine and hypoxanthine were separated from platelet-poor plasma.After thrombin treatment, radioactive ATP in platelets broke down rapidly, while radioactive ADP decreased more slowly. Radioactive AMP increased at first in the cellular and supernatant fractions, and then decreased gradually. The accumulation of radioactive hypoxanthine was observed in the supernatant fraction. Released radioactive ATP and ADP were 23% and 22% of the initial radioactive ATP and ADP in platelets, respectively.

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.

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.

Effects of ATP precursors on ATP and free ADP content and functional recovery of postischemic hearts

1989 ◽  
Vol 256 (2) ◽  
pp. H560-H566 ◽  
Author(s):  
G. Ambrosio ◽  
W. E. Jacobus ◽  
M. C. Mitchell ◽  
M. R. Litt ◽  
L. C. Becker
Keyword(s):  
Functional Recovery ◽  
Adenine Nucleotide ◽  
Mean Value ◽  
Global Ischemia ◽  
Stunned Myocardium ◽  
Base Line ◽  
Atp Content ◽  
Atp Levels ◽  
Developed Pressure

It has been proposed that administration of adenine nucleotide precursors might accelerate replenishment of myocardial ATP and "free" ADP, thus improving recovery of depressed contractility of postischemic hearts. To test this hypothesis, Langendorff-perfused rabbit hearts were subjected to 20 min of global ischemia and reperfused for 2 h with normal perfusate (n = 8) or perfusate containing 100 mumol/l of the ATP precursors adenosine (n = 8) or 5-amino-4-imidazolecarboxamide riboside (AICAriboside; n = 8). After reperfusion, developed pressure in untreated hearts averaged 70-80% of base line, whereas ATP content was reduced to approximately 70% of preischemic values. AICAriboside administration did not increase tissue ATP levels or contractility. However, in every heart that received adenosine during reperfusion, ATP content increased from a mean value of 65 +/- 4% of base line to 84 +/- 5% at the end of reperfusion (P less than 0.001). Free ADP also increased in adenosine-treated hearts from 40 to 50% of base line at the beginning of reperfusion, to normal levels by 60 min. However, no improvement in contractility was observed in the hearts that received adenosine. These results support the hypothesis that decreased availability of nucleotide precursors is responsible for depressed ATP levels in postischemic hearts; however, reduced ATP and free ADP levels may not be directly responsible for the depressed function of stunned myocardium.

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 Initial Changes in the Level of Adenine Nucleotides By Glucose Addition in Ehrlich Ascites Tumor Cells.

1963 ◽  
Vol 17 ◽  
pp. 881-882 ◽  
Author(s):  
Kay Overgaard-Hansen ◽  
Lars Ernster ◽  
Rolf Luft ◽  
Lars Ernster ◽  
Jon Munch-Petersen
Keyword(s):  
Tumor Cells ◽  
Adenine Nucleotides ◽  
Ehrlich Ascites Tumor ◽  
Ehrlich Ascites Tumor Cells ◽  
Ascites Tumor ◽  
Glucose Addition ◽  
Ehrlich Ascites

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.

Contribution from local conversion of thyroxine to 3,5,3'-triiodothyronine to intracellular 3,5,3'-triiodothyronine in several organs in hypothyroid rats at isotope equilibrium

1982 ◽  
Vol 101 (3) ◽  
pp. 386-396 ◽  
Author(s):  
J. van Doom ◽  
F. Roelfsema ◽  
D. van der Heide
Keyword(s):  
Cerebral Cortex ◽  
Pituitary Gland ◽  
Subcellular Fractions ◽  
Thigh Muscle ◽  
Intracellular Level ◽  
Carrier Free ◽  
Tissue Homogenates ◽  
Isotope Equilibrium ◽  
Layer Chromatography ◽  
The Brain

Abstract. The intracellular conversion of T4 to T3 was investigated in various tissues of hypothyroid rats after continuous iv infusion of radiolabelled T3 and T4. Two groups of 4 thyroidectomized rats were infused with carrier-free 125I-labelled T4 as well as 131I-labelled T3 until isotope equilibrium was achieved. Plasma, various tissue homogenates (liver, kidney, pituitary, thigh muscle, cerebral cortex and cerebellum) and subcellular fractions (nuclei, mitochondria, microsomes, cytoplasm) from liver, kidney and the pituitary gland were extracted for thin layer chromatography. The [125I]T3/[131I]T3 ratios were determined and the extra contribution of [125I]T3 derived from local conversion of [125I]T4 to the total [125I]T3 was calculated in percent. In addition to the [125I]T3 derived from plasma, [125I]T3 derived from locally converted [125I]T4 was present in all tissues investigated. There was substantially more, although in varying quantities, in the cerebral cortex (79 ± 2%), the cerebellum (68 ± 4%) and the pituitary gland (53 ± 1%) than in the liver (10 ± 6%), the kidney (11 ± 5%) and thigh muscle (17 ± 6%); in the latter tissues most of the 125I-labelled T3 is derived directly from plasma. These results indicate that in the brain of severe hypothyroid rats there is pronounced conversion of T4 to T3 and effective binding of the T3 produced whereas the T3 in the liver, kidney, and muscle is predominantly derived from plasma. At the intracellular level, within the investigated tissues, the locally formed T3 was distributed equally over the subcellular fractions.

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