Regulation of active Na+-K+ transport in skeletal muscle.
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Adrenalectomy↗
Biomedical subjects
T Clausen
Publications and source records attributed to T Clausen.
[The Na,K pump, the cellular receptor for digitalis glycosides].
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Cell Membrane↗
Effects of ouabain, age and K-depletion on K-uptake in rat soleus muscle.
The relationship between the number of 3H-ouabain binding sites and the Na, K-pump mediated K-uptake has been characterized in rat soleus muscle. By brief exposure to 3H-ouabain (1 X 10(-6)-1 X 10(-5) mol/l) in vitro, it could be measured that 19-94% of the ouabain binding sites had been occupied. This was associated with a proportionate decrease in the ouabain suppressible K-uptake indicating that under strictly standardized conditions, measurements of 3H-ouabain binding sites quantify functional Na,K-pumps. When 3 week old rats were K-depleted for a further week followed by K-repletion 2 h before measurements, the 3H-ouabain binding site concentration was 61% lower than in age-matched control soleus muscles. However, the ouabain suppressible K-uptake was only reduced by 35%, partly because intracellular Na remained higher in the muscles obtained from K-depleted rats. From the 1st to the 4th week of life, the 3H-ouabain binding site concentration increased 2.9-fold. In contrast, the ouabain suppressible K-uptake decreased by a factor 3.5. Accordingly, in muscles from 1 week old rats, the ouabain suppressible K-uptake per 3H-ouabain binding site was 10-fold higher than in muscles from 4 week old rats. This difference could not be accounted for by changes in intracellular Na, total or extracellular water. It may be related to differentiation and change in structure.(ABSTRACT TRUNCATED AT 250 WORDS)
Aging↗
The significance of the effects of insulin on Na,K-transport in muscle cells.
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Amino Acids↗
Significance of skeletal muscle digitalis receptors for [3H]ouabain distribution in the guinea pig.
The importance of specific digitalis glycoside binding sites in skeletal muscle for the digitalis glycoside distribution in the guinea pig was evaluated using [3H]ouabain and [3H]digoxin binding assays. Measurements of [3H]ouabain binding capacity (EOmax) in gastrocnemius and heart muscles in vitro gave values of 474 +/- 15 and 1,092 +/- 39 pmol/g wet wt., respectively, in 4-week-old guinea pigs. Hence the total amount of [3H]ouabain binding sites in skeletal muscle and the heart was around 42,700 and 1,200 pmol, respectively. The apparent dissociation constants (Kd) for ouabain receptor interaction was 0.7 X 10(-7) and 1.5 X 10(-7) M for skeletal muscle and heart, respectively. Comparison of [3H]ouabain and [3H]digoxin binding revealed that these drugs are competitive. From birth to maturity the concentration of [3H]ouabain binding sites in guinea pigs decreased from 803 +/- 58 to 304 +/- 28 pmol/g wet wt. in gastrocnemius muscle and from 1,458 +/- 31 to 1,079 +/- 19 pmol/g wet wt. in the heart. After i.p. injection, measurements of the distribution of [3H]ouabain in plasma, skeletal muscle and the heart showed an almost equal relative specific occupancy of digitalis glycoside receptors in skeletal muscle and the heart: When 10% of the digitalis receptors in the heart were occupied by [3H]ouabain, 13% of those in the skeletal muscles were occupied. It was calculated that 1 hr after the i.p. administration of [3H]ouabain the amount of [3H]ouabain specifically bound to the skeletal muscles and the heart corresponded to 5 times and 1/10 the amount available in the extracellular pool, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
Age Factors↗
Effect of thyroid function on number of Na-K pumps in human skeletal muscle.
To evaluate the effect of thyroid function on the number of Na-K pumps in skeletal muscle, the number of 3H-ouabain binding sites was measured in biopsy specimens from the vastus lateralis muscle of euthyroid subjects and patients with hypothyroidism or hyperthyroidism. In hypothyroidism there was a decrease of 50% in 3H-ouabain binding sites, and in hyperthyroidism there was an increase of 68% in 3H-ouabain binding sites. When thyroid status became normal after treatment the number of 3H-ouabain binding sites also became normal. There were significant correlations between several thyroid function tests and the number of 3H-ouabain binding sites. Free T4-index gave the highest r value (0.87). These changes may account for the variations in digitalis sensitivity associated with thyroid disorders.
Adult↗
Energetics of active sodium-potassium transport following stimulation with insulin, adrenaline or salbutamol in rat soleus muscle.
The total metabolic energy expenditure associated with active Na-K-transport over the first 20 min of stimulation with insulin, adrenaline or salbutamol (delta HmNa-K) was determined from direct calorimetric and tracer ion flux measurements in isolated muscles at rest. The reversible work performed by the Na-K-pump during the same interval of time (WrevNa-K- was calculated as the product of the ouabain-suppressible Na-K transfers and the mean free energy increase imparted to the two ions as they are transported against their electrochemical gradients across the plasma membrane. Comparison of membrane potential and intracellular Na and K concentrations before and after the stimulation indicated that part of WrevNa-K had contributed to increase in ion electrochemical gradients in the preparation (i.e. had not been lost as heat) during the 20 min period. Accordingly, the maximum value of delta HmNa-K was taken as the sum of the ouabain-suppressible heat production and WrevNa-K. Following stimulation with insulin, adrenaline or salbutamol this maximum corresponded to 10, 10 and 12% respectively, of basal metabolism. Under the same three conditions, the minimum "energetic efficiency" of the active Na-K-transport process, defined as the ratio between WrevNa-K and maximum delta HmNa-K, was 35, 41 and 38%, respectively.
Albuterol↗
The age-dependent changes in the number of 3H-ouabain binding sites in mammalian skeletal muscle.
The influence of age on the binding of 3H-ouabain in skeletal muscle has been characterized in rats, mice and guinea pigs. Measurements performed using biopsies and intact fibers obtained from different types of rat muscles showed that from birth to the 4th week of life, the number of 3H-ouabain binding sites per unit weight increases up to 5-fold, followed by almost the same relative decrease to a plateau around 250 pmol/g wet wt at an age of 22 weeks. These changes were not associated with any major alterations in apparent KD (1.7-3.1 X 10(-7) M) dissociation rate or heterogeneity in binding characteristics. Measurements of 3-O-methylfluorescein phosphatase activity, an enzyme activity which is closely correlated to the Na-K-ATPase activity, confirmed the 3H-ouabain binding data. In mice, the number of 3H-ouabain binding sites showed similar, albeit less pronounced changes with age, a maximum being reached at the 4th week of life. In guinea pigs, the number of 3H-ouabain binding sites per unit weight decreased by 60% from birth to maturity. The results indicate that the early development and differentiation of individual skeletal muscles is associated with a marked increase in the number of Na-K-pumps (when expressed as pmol/muscle), until at maturity a plateau is reached. However, when expressed as pmol/g wet wt the increase is followed by a decrease to a plateau. This may in part account for the relatively low digitalis sensitivity seen in infants as compared to newborn and mature individuals.
Aging↗
Effect of K-depletion on 3H-ouabain binding and Na-K-contents in mammalian skeletal muscle.
K-depletion induced by K-deficient fodder led to hypokalemia, decreased K-content and increased Na-content in skeletal muscle of rats. The heart showed similar, although more modest changes, whereas brain, erythrocytes and liver maintained virtually constant Na-K-contents. K-depletion decreased total binding capacity for 3H-ouabain by up to 76%, an effect which could be demonstrated both in vitro and in vivo. Following 3 weeks of K-depletion, the apparent KD for 3H-ouabain binding to rat soleus was 1.3 X 10(-7) M as compared to 2.4 X 10(-7) M in controls. Also in mice and guinea pigs, K-depletion induced a selective loss of K from muscle and decreased 3H-ouabain binding capacity. K-depletion induced by diuretics or fluorohydrocortisone gave similar effects. The effects of K-depletion on 3H-ouabain binding capacity were confirmed by measurements of 3-O-methylfluorescein phosphatase activity, an enzyme activity which is closely correlated to the Na-K-ATPase activity. Following readministration of K, the K-contents of plasma and muscle reached control levels in 24 hours, but 3H-ouabain binding capacity was not normalized until after 6 days of K-repletion. In mammalian skeletal muscle, K-depletion leads to a marked and reversible reduction in 3H-ouabain binding capacity, which may be secondary to the selective loss of K or gain of Na.
Animals↗
Effect of thyroid status on K+-stimulated metabolism and 45Ca exchange in rat skeletal muscle.
In the perfused hindlimbs of hypo-, eu-, and hyperthyroid rats, high K+o (20 mM) markedly stimulated glucose uptake, lactate production, and O2-consumption to levels increasing with the thyroid status. At Ca2o+ less than 50 microM, all the responses became transient. In the hypothyroid preparations, caffeine (5 mM) produced no stimulation, but in those obtained from eu- and hyperthyroid rats, it increased all the metabolic parameters as well as K+ release in proportion to thyroid status. In isolated soleus and extensor digitorum longus muscles, both high K+o and caffeine stimulated 45Ca-efflux, with the response increasing in proportion to thyroid status. This stimulation was reduced by 70-100% with dantrolene (10(-5) M), which in earlier studies was shown to suppress the metabolic effects of high K+o and caffeine. High K+o also increased 45Ca-influx but produced no change in muscle calcium contents. The results support the idea that thyroid hormones control substrates availability and energy consumption in skeletal muscle by increasing the amount of sarcoplasmic reticulum and thereby the rate of Ca2+ mobilization into the cytosol.
Animals↗
A method for the determination of the total number of 3H-ouabain binding sites in biopsies of human skeletal muscle.
A new method based on vanadate facilitated binding of 3H-ouabain has been applied for the quantitative determination of the number of 3H-ouabain binding sites (Na-K-pumps) in needle biopsies of human skeletal muscle. Samples of the vastus lateralis muscle weighing 2-8 mg showed specific and saturable binding of 3H-ouabain with an apparent KD of 1.9 X 10(-8) mol/l. In 20 healthy human subjects in the age range 25-80 years, the number of 3H-ouabain binding sites was 278 +/- 15 pmol/g wet weight with no relation to age or sex. In samples of the intercostal and rectus abdominis muscles, the number of 3H-ouabain binding sites varied from 225 to 280 pmol/g wet weight. These values are at least 2 times higher than those previously reported for human skeletal muscle. The number of 3H-ouabain and 3H-digoxin binding sites were identical, and ouabain (10(-3) mol/l) completely displaced specifically bound 3H-digoxin. When biopsies were frozen in liquid N2 immediately after withdrawal, storage at -20 degrees C for up to 11 weeks caused no significant change in the number of 3H-ouabain binding sites. The method allows quantitative determination of the number of 3H-ouabain binding sites in standard biopsies of human skeletal muscle to be performed by simple procedures within a few hours. This can be used for the study of conditions where the number of Na-K-pumps is known to undergo fluctuations.
Adult↗
A simple and rapid method for the determination of the number of 3H-ouabain binding sites in biopsies of skeletal muscle.
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Animals↗
Adrenergic control of Na+-K+-homoeostasis.
Catecholamines induce hypokalaemia via stimulation of beta-adrenoceptors, primarily in skeletal muscle but also in other tissues. This is the result of increased active Na+-K+-transport, leading to a rise in the intracellular K+/Na+-ratio and hyperpolarisation in muscle cells. These effects are mediated by 3', 5'-cyclic adenosine monophosphate, can be detected down to physiological concentrations of adrenaline and noradrenaline, and are seen both in vitro and in vivo. Catecholamines released from the adrenal medulla as well as sympathetic nerve endings are of importance in clearing K+ from the extracellular water space during K+-loading or exercise. beta 2-adrenoceptor agonists can be used in the treatment of hyperkalaemia, and beta-adrenoceptor blockade may induce hyperkalaemia, in particular during exercise. The effects of catecholamines on the contractile performance of skeletal muscles are partly due to stimulation of the active electrogenic Na+-K+-transport across the sarcolemma.
Adrenergic beta-Agonists↗
Effects of denervation on sodium, potassium and [3H]ouabain binding in muscles of normal and potassium-depleted rats.
K depletion leads to a selective loss of K from skeletal muscles, which is associated with a decrease in the number of [3H]ouabain binding sites. The significance of the nerve supply for these changes has been assessed in denervation experiments with K-depleted rats. In K-depleted rats (age 4-12 weeks) denervation led to a partial recovery of the K contents in soleus (46-77%), gastrocnemius (23%) and extensor digitorum longus (e.d.l.) muscles (19%) within 24 h. These effects were not prevented by beta-adrenoceptor blockade or mimicked by alpha-adrenoceptor blockade. In K-depleted rats the number of [3H]ouabain binding sites was not increased following denervation. In K-depleted rats 24 h of plaster immobilization of the entire hind limb caused 51% recovery of the total K content in soleus, whereas gastrocnemius and e.d.l. showed 49 and 16% recovery, respectively. Tenotomy for 3 h caused a rise in total K content of 33% in soleus muscles from K-depleted rats. Anaesthesia for 3 h increased the total K content by 23%. The recovery of K induced by denervation, immobilization in plaster, tenotomy or anaesthesia was associated with an equivalent decrease in Na content. Denervation performed before K depletion reduced the loss of K from soleus, but not from gastrocnemius and e.d.l. In both soleus and e.d.l. the number of [3H]ouabain binding sites, however, decreased to the same level as in the contralateral innervated muscles. Denervation reduced, but did not prevent, the increase in the number of [3H]ouabain binding sites seen after re-administration of K to K-depleted rats. It is concluded that the changes in Na-K contents seen after denervation in K-depleted rats are the outcome of cessation of muscle activity. The results give no support to the idea that the effects of K depletion on the K content and the number of [3H]ouabain binding sites in skeletal muscle are mediated by the peripheral nerves.
Animals↗
The stimulating effect of 3',5'-(cyclic)adenosine monophosphate and lipolytic hormones on 3-O-methylglucose transport and 45Ca2+ release in adipocytes and skeletal muscle of the rat.
(1) In order to assess the possible role of 3',5'-(cyclic)adenosine monophosphate (cAMP) in the control of glucose transport, the effect of the nucleotide or agents known to increase its intracellular concentration on sugar transport or 45Ca2+ washout were characterized in epididymal fat pads, free fat cells and soleus muscles of the rat. (2) When added to the incubation medium, cAMP (0.1-2.0 mM) stimulated 3-O-[14C]methylglucose washout from fat pads. This effect was abolished by cytochalasin B, and additive to that induced by submaximal (10-25 microU/ml), but not by supramaximal (10 microU/ml) concentrations of insulin. (3) cAMP (2 mM) stimulated the conversion of [U-14C]glucose into CO2 and triacylglycerols. This effect was additive to that of insulin (100 microU/ml). (4) ACTH, glucagon, adrenaline, noradrenaline and salbutamol, which are all known to increase the cAMP content of adipose tissue, stimulated the washout of 3-O-[14C]methylglucose and 45Ca2+ from preloaded fat pads. The fractional losses of the two isotopes were significantly correlated (P less than 0.001, r = 0.73). (5) In free fat cells, adrenaline (10(-6) M) and salbutamol (10(-5) M) stimulated the uptake of 3-O-[14C]methylglucose, and salbutamol (10(-5) M) did not interfere with the stimulating effect of insulin (25 microU/ml) on sugar uptake. (6) In rat soleus muscles, adrenaline and salbutamol produced a dose-dependent stimulation of the washout of 3-O-[14C]methylglucose and 45Ca2+. The effect of adrenaline on sugar efflux was abolished by propranolol. (7) It is concluded that the activation of the glucose transport system by insulin is unlikely to be mediated by a drop in the cellular concentration of cAMP. An increase in cAMP brought about by beta-adrenoceptor agonists or lipolytic hormones may induce a mobilization of calcium ions from cellular pools into the cytoplasm, which in turn leads to the activation of the glucose transport system demonstrated in the present as well as in several earlier studies.
3-O-Methylglucose↗
Age-dependent changes in the number of [3H]ouabain-binding sites in rat soleus muscle.
The influence of age on the number of (Na+K+)-ATPase units in skeletal muscle has been assessed by measurements of [3H]ouabain binding in vitro and in vivo to rat soleus muscle. In vitro measurements showed that from the 2nd to the 28th day of life, the number of [3H]ouabain-binding sites increases from 120 to 580 pmol/g wet wt. This is followed by a decrease, until a plateau between 150 and 200 pmol/g is reached around 150 days after birth. 60 min after intraperitoneal injection of [3H]ouabain (12.5 mumol/kg body weight), the soleus muscles of 28-day-old rats had accumulated 2.4-times more 3H-activity per g wet wt. than muscles of 85-day-old rats and the 3H-activity in plasma was 54% lower. The results may explain the low sensitivity to digitalis glycosides found in infants as compared to premature or adult individuals.
Aging↗
The Na+-K+-pump, energy metabolism, and obesity.
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Animals↗
Desensitization by terbutaline of beta-adrenoceptors in the guinea-pig soleus muscle: biochemical alterations associated with functional changes.
1 The effects of adrenaline and terbutaline on cyclic adenosine 3',5'-monophosphate (cyclic AMP) content, 22Na-efflux, 42K-influx and subtetanic contractions have been assessed in soleus muscles isolated from guniea-pigs which had been maintained on food with or without terbutaline for 5 days. 2 Terbutaline and adrenaline increased cyclic AMP content and suppressed subtetanic contractions, and regression analysis indicates a statistically significant correlation between these two effects (P less than 0.01). 3 In muscles obtained from terbutaline-treated animals, the effects of terbutaline and adrenaline on cyclic AMP content, active Na-K-transport and subtetanic contractions were all considerably suppressed, but insulin stimulated 22Na-efflux and affected subtetanic contractions to the same extent as in the muscles obtained from the control group. 4 The results suggest that terbutaline treatment leads to a reduction in the number of beta 2-adrenoceptors in skeletal muscle or an impairment of their function. 5 The results provide further support for the idea that the effect of adrenaline or insulin on skeletal muscle contractions is the outcome of stimulation of active Na-K-transport.
Animals↗