THE SELECTIVE ABSORPTION OF POTASSIUM BY ANIMAL CELLS : III. THE EFFECT OF HYDROGEN ION CONCENTRATION UPON THE RETENTION OF POTASSIUM.

By perfusing frogs for varying periods with potassium-free Ringer solutions having a pH ranging from 6.0 to 8.0, it has been determined that such solutions have little or no effect upon the retention of potassium by muscle cells.     

THE SELECTIVE ABSORPTION OF POTASSIUM BY ANIMAL CELLS : II. THE CAUSE OF POTASSIUM SELECTION AS INDICATED BY THE ABSORPTION OF RUBIDIUM AND CESIUM.

1. Frog muscles perfused with Ringer solution in which potassium chloride has been replaced by an equivalent amount of rubidium or cesium chloride take up rubidium or cesium and incorporate them into the tissue substance in such form as to be retained during a subsequent perfusion with potassium-free Ringer solution, provided the muscles contract during the first perfusion. Retention of rubidium or cesium by a resting muscle does not occur. 2. Rats on synthetic diets, adequate in all respects except that potassium was replaced by an equivalent amount of rubidium or cesium, died after a period varying from 10 to 17 days with characteristic symptoms including tetanic spasms. Muscle, heart, liver, kidney, spleen, and lung tissues were then found to contain significant amounts of rubidium or cesium. The concentration of these metals in the muscle amounted, in some cases, as shown by a spectroscopic estimation, to about half the concentration of potassium normally found in mammallian muscle. 3. The results are regarded as tending to confirm the theory that the peculiarities in the physiological effects of potassium, including the facility with which it is "selected" by living cells in preference to sodium, are related to the electronic structure of the potassium ion as compared with that of similar ions. The possible relationship of the comparative migration velocity, a function of the electronic structure, to physiological effects is suggested.     

THE POTASSIUM EQUILIBRIUM IN MUSCLE

1. Analyses were made of the K and HCO₃ content, the irritability, and weight change of isolated frog sartorius muscles after immersion for 5 hours in Ringer''s solutions modified as to pH and potassium content. 2. At each pH a concentration of potassium in the solution was found which was in diffusion equilibrium with the potassium in the muscle. In greater concentrations potassium moved into the muscle against the concentration gradient and vice versa. 3. The greater the alkalinity of the solution the smaller the concentration of the potassium at equilibrium so that the product of the concentrations of OH and K in the solution at equilibrium tends to remain approximately constant. 4. The pH inside the muscle is approximately equal to that outside when first dissected but it tends to change during immersion so as to follow the changes in the pH of the solution. This finding is in direct conflict with the theory according to which the high potassium concentration inside should be accompanied by an equally high hydrogen ion concentration in relation to that outside. 5. The diffusion of potassium into the muscle makes its contents more alkaline but the increase in alkalinity is not always, nor usually, equivalent to the amount of potassium which has diffused and conversely, the pH inside can change in either direction according to the pH outside without there being any diffusion of potassium. Hence potassium is not the only penetrating ion. 6. The irritability of the muscles is at a maximum in concentrations of potassium which are greater than that in normal Ringer''s solution, or about 20 mg. per cent potassium. This optimum does not seem to be a function of pH and is therefore not dependent upon the direction of movement of the potassium but probably on the ratio of potassium outside to that inside. 7. Swelling of the muscles occurs in solutions which injure the muscle so as to permit both cations and anions to enter without permitting the organic protein anions to escape. Anion impermeability is necessary to prevent this same osmotic swelling under normal conditions. 8. An increase in the CO₂ tension in muscle and solution causes a greater increase in acidity in the solution than in the muscle and leads to a loss of potassium. One expects therefore a potassium shift from tissues to blood comparable to the chlorine shift from plasma to corpuscles.     

空心莲子草、大豆和向日葵根部K~+(~(86)Rb~+)的吸收和通量分析

与大豆、向日葵相比,空心莲子草根系具有一个较高效率的K~+吸收机制。它的K_m值很小,I_m值又较高。空心莲子草根系将K~+向地上部运输的能力也比大豆和向日葵高得多。 上述三种植物根组织K~+含量的差异主要是因为液泡中含K~+量的不同,而细胞质的含K~+量影响不大。其中,参数K_cv/K_vc的比值可以表示液泡积累K~+的特征,并且也决定着不同植物根组织积累K~+的能力。     

Different modes of activating phosphofructokinase,a key regulatory enzyme of glycolysis,in working vertebrate muscle

Glycolytic flux in white muscle can be increased several-hundredfold by exercise. Phosphofructokinase (PFK; EC 2.7.1.11) is a key regulatory enzyme of glycolysis, but how its activity in muscle is controlled is not fully understood. In order not to neglect integrative aspects of metabolic regulation, we have studied in frogs (Rana temporaria) a physiological form of muscle work (swimming) that can be triggered like a reflex. We analysed swimming to fatigue in well rested frogs, recovery from exercise, and repeated exercise after 2 h of recovery. At various times, gastrocnemius muscles were tested for glycolytic intermediates and effectors of PFK. All metabolites responded similarly to the two periods of exercise, with the notable exception of fructose 2,6-bisphosphate (F2,6P(2)), which we proved to be a most potent activator of frog muscle PFK. The first bout of exercise triggered a more than 10-fold increase in F2,6P(2); PFK activity and the content of F2,6P(2) in muscle were well correlated. F2,6P(2) decreased to pre-exercise levels in fatigued frogs and it virtually disappeared during recovery. Varying by a factor of 70, F2,6P(2) was the most dynamic of all metabolites in muscle. Even more surprisingly, F2,6P(2) did not respond at all to a second bout of exercise. Other activators of PFK, such as Pi, AMP and ADP, are increased as a consequence of increased ATP turnover in contracting muscle cells. This does not apply to F2,6P(2) which is likely to respond to extracellular signals and could be involved in mechanisms by which muscle metabolism is integrated into the metabolism of the whole body. Whether this phenomenon exists in vertebrates other than the frog, and maybe even in humans, and how the content of F2,6P(2)in muscle is controlled are intriguing open questions.     

Ryanodine modification of cardiac muscle responses to potassium-free solutions. Evidence for inhibition of sarcoplasmic reticulum calcium release

To test whether ryanodine blocks the release of calcium from the sarcoplasmic reticulum in cardiac muscle, we examined its effects on the aftercontractions and transient depolarizations or transient inward currents developed by guinea pig papillary muscles and voltage-clamped calf cardiac Purkinje fibers in potassium-free solutions. Ryanodine (0.1-1.0 microM) abolished or prevented aftercontractions and transient depolarizations by the papillary muscles without affecting any of the other sequelae of potassium removal. In the presence of 4.7 mM potassium and at a stimulation rate of 1 Hz, ryanodine had only a small variable effect on papillary muscle force development and action potential characteristics. In calf Purkinje fibers, ryanodine (1 nM-1 microM) completely blocked the aftercontractions and transient inward currents without altering the steady state current-voltage relationship. Ryanodine also abolished the twitch in potassium-free solutions, but it enhanced the tonic force during depolarizing voltage- clamp steps. This latter effect was dependent on the combination of ryanodine and potassium-free solutions. The slow inward current was not blocked by 1 microM ryanodine, but ryanodine did appear to abolish an outward current that remained in the presence of 0.5 mM 4- aminopyridine. Our observations are consistent with the hypothesis that ryanodine, by inhibiting the release of calcium from the sarcoplasmic reticulum, prevents the oscillations in intracellular calcium that activate the transient inward currents and aftercontractions associated with calcium overload states.     

Nonquantum release of acetylcholine in frog neuromuscular junction after blocking of axoplasmic transport with colchicine

Standard microelectrode techniques were used to evaluate the effect of d-tubocurarine chloride on membrane potential of junctional and extrajunctional areas of muscle fibers in a potassium-free Ringer solution. The experiments were made on frogs after inactivation of acetylcholinesterase. d-Tubocurarine chloride hyperpolarized the membrane of muscle fibers only in the junctional area. Blockade of axoplasmic transport with colchicine did not affect the magnitude of the hyperpolarization response of the membrane end plate to the presence of d-tubocurarine chloride, but at the same time it significantly reduced the membrane rest potential of muscle fibers, and gave rise to the appearance of extrajunctional sensitivity to acetylcholine. It is concluded that the blockade of axoplasmic transport does not affect the pattern of non-quantum acetylcholine release from nerve terminals. Therefore, this is unlikely to cause denervation-like changes in the muscle under the conditions described.     

Estimation of the levels of calcium and other electrolytes during various phases of contraction of the longitudinal smooth muscle of the guinea pig ileum

A method is described for measuring calcium and other electrolyte levels during various phases of contraction of the longitudinal smooth muscle of the guinea pig ileum. Muscles are immersed in a solution of 160 mM Tris-Cl containing 10 mM lanthanum for 30 min to reduce the contribution of extracellularly bound cations, while the temperature is reduced to 4°C to prevent active cation fluxes. The muscle cells gained calcium and lost magnesium during the phasic and tonic phases of contractions by the muscarinic agent, CD (cis-2-methyl-4-dimethylaminomethyl-1, 3-dioxolane methiodide) and during the tonic phase of contractions by KCl. Muscles lost potassium during contractions to CD, but gained potassium during contractions to 60 mM KCl. The potassium lost during contractions to CD was regained slowly over a period of 30 min when the CD was washed out. This recovery corresponded to the rate of recovery of spontaneous activity and normal responsiveness of the muscle to further doses of stimulants. Calcium levels were reduced rapidly to below normal levels on washout of the CD and were regained in a similar time course to that of potassium. Sodium levels were not significantly changed with either stimulant.     

Generation of nonadrenergic inhibitory junction potentials in the smooth muscle of the guinea-pig gastrointestinal tract after replacing potassium ions with cesium ions

Nonadrenergic inhibitory junction potentials (IJPs), evoked by intramural nerve stimulation, were studied in the smooth muscle of the guinea-pig stomach, cecum, and colon, using a modified sucrose-gap technique. After incubating smooth muscle preparations for 4–9 h in potassium-free Krebs solution, IJPs were abolished, but reappeared when cesium ions (6 mM) were added to the Krebs solution. Under these conditions, in the majority of cases the amplitude of the IJP was half as small, and the latency and duration were significantly longer, than in normal conditions; also ATP, but not adenosine, caused hyperpolarization of the smooth muscle membrane. The amplitude of the IJP depended on the extracellular concentration of cesium. In all types of preparation, in cesium-containing Krebs solution, apamin usually abolished the IJP and responses to ATP. These results are consonant with the purinergic hypothesis of inhibitory neuromuscular transmission. The generation of the IJP in these potassium-free conditions depends on cesium ions, which pass through the small-conductance apamin-sensitive, calcium-dependent potassium channels.A. A. Bogomoletz Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 22, No. 5, pp. 634–641, September–October, 1990.     

不同供钾水平对小麦旗叶光合速率日变化的影响

在田间栽培条件下,研究了不同供钾水平对小麦(Triticum aestivum)‘安农8903’旗叶光合速率日变化的影响。结果表明：从扬花期到灌浆中期（5月1～20日）,植株及叶片中钾含量随不同供钾水平的提高而增大,尤以施K 82.5 kg·hm-2的增幅较大（2.7倍）。在小麦籽粒的灌浆进程中,植株的含钾量不断增加,但叶片中的含钾量却呈下降趋势。扬花期到灌浆中期的小麦旗叶由于高光强、高温和较低的相对湿度,其气孔导度和净光合速率日变化均呈双峰曲线。两者皆在10∶00左右和15∶00左右各有一个峰值,前一峰值高于后一峰值,两峰之间的低谷在14∶00左右。胞间CO2浓度日变化则呈相反趋势,两个低谷出现于12∶00和15∶00,但灌浆中期时日变化曲线中只有一个低谷。土壤增施钾肥可增大小麦旗叶的气孔导度和净光合速率,减轻10%～37%由光合午休造成的CO2同化量损失,提高小麦产量。但施钾量与胞间CO2浓度之间相关性不显著。4种不同的钾素水平处理中以施用K2O 165 kg·hm-2的效果最佳。     

The effect of ions upon the response of smooth muscle to cooling

The slow tonic responses of the anterior byssus retractor of Mytilus edulis to rapid cooling were investigated by simultaneously recording tension and resting potential changes after soaking the muscle in banthine, a powerful neuromuscular blocking agent. The quantitative relations between the amount of cooling and the amount of associated depolarization necessary for contraction at various concentrations of potentiating potassium can be expressed in a family of curves. The plateaus of the curves for sea water and for potassium-free sea water were beneath the depolarization value necessary for contraction, so that it is clear that no amount of cooling with sea water alone or with potassium-free sea water would ever be effective. When the muscle is treated with subthreshold amounts of potassium and rapidly cooled in various concentrations of sodium ion and calcium ion, respectively, the sodium and calcium do not affect the amount of depolarization. Acetylcholine, in subthreshold amounts, has a potentiating effect, but, unlike potassium and cooling, acts through the nervous apparatus. Mytilus muscle will respond to cooling with tonic contraction whenever a critical threshold amount of depolarization is achieved. Cooling alone cannot trigger the contraction since it cannot bring about sufficient depolarization. Cooling can result in contraction, however, if used in conjunction with some other subthreshold depolarizing agent. Cooling affects the contractile mechanism by first causing membrane breakdown and depolarization.     

EFFECTS OF POTASSIUM AND LITHIUM ON SODIUM TRANSPORT FROM BLOOD TO CEREBROSPINAL FLUID

Abstract— The relationship between isotopic sodium entry into cerebrospinal fluid (CSF) from blood and cisternal potassium concentration was studied using ventriculo-cisternal perfusion in the rabbit. The entry of sodium into CSF was separated into 2 components. The fast component was significantly correlated with cisternal potassium concentration during perfusions with a potassium-free artificial CSF. ATPase activity in the homogenised choroid plexus was shown to be sensitive to potassium over a range of concentrations similar to that in the perfusion studies. The results are interpreted as showing a potassium-sensitive entry of ²⁴Na across the choroid plexus due to a sodium-pump situated in the apical membrane of the choroid plexus. The effects of low concentrations of lithium (0.6–1.2 mm) on ²⁴Na entry into CSF and brain and on CSF secretion were studied. When applied via the ventricles lithium caused a 30–39% stimulation of the fast component of sodium entry and a 28% stimulation of CSF secretion. When given via the blood lithium inhibited the fast component of sodium entry and CSF secretion by 43% and 40% respectively. No effects of lithium were found on the slow component of sodium entry into CSF or sodium entry into brain. The results suggest that lithium at low (0.6–1.2 mM) concentrations can stimulate the choroid plexus sodium-pump at the potassium-sensitive side and inhibit it at the sodium-sensitive side.     

The effects of modified ringer's solutions on the membrane potentials of innervated and denervated frog sartorius muscle fibers

Comparison has been made between innervated and chronically denervated frog sartorius muscle fibers for resting potentials and a number of features of the action potential. Muscles were obtained from force-fed frogs maintained at room temperature for periods up to one year, and were studied with intracellular microelectrodes. Denervated muscles increased in sensitivity to acetylcholine by 100–400-fold. Studies were made in normal Ringer's solution, and in media in which concentrations of K⁺, Na⁺, Ca⁺⁺, and Cl? were altered. The only significant differences noted between the denervated and the innervated fibers were a reduction in the maximum rate of fall of the action potential (ca. 20%) and an increase in the fall time of the active membrane potential (ca. 25%). These differences were present in normal Ringer's solution and remained when the bathing medium was modified. The resting membrane potential of denervated and innervated muscles varied with log [K⁺]_o in exactly the same manner, and followed the theoretical relation proposed by Hodgkin (Proc. Roy. Soc., B, 148: 1–37, ′58), with the term representing the ratio of the sodium to potassium permeabilities assigned a value of 0.01. The results suggest that (a) the resting sodium and potassium permeabilities are reduced proportionately after denervation, since it is known that denervated frog muscle has a smaller potassium permeability, and (b) the mechanism controlling the increase in potassium conductance during the action potential is less available after denervation. Data indicate that the system controlling the sodium permeability is capable of activation to the same extent as in innervated muscles. Muslces which had been allowed to reinnervate did not show the differences presented by the denervated muscles. Innervated and denervated muscles did not show any significant changes in maximum rates of rise or fall of the action potential, nor of the active membrane potential amplitude over a 30 mV range of resting membrane potentials, indicating that the sodium and potassium permeability systems are fully available in frog muscle at membrane potentials larger than ?80 mV.     

Oxygen consumptions and potassium contents of slices of rat renal cortex.

An oxygen electrode respirometer for determining the oxygen consumption of slices of mammalian renal cortex is described and assessed. Though rat renal cortical slices incubated in potassium-free medium for one hour lost 102 +/- 14 mmol of potassium/kg dry weight, there was only a small, nonsignificant fall in oxygen consumption. In contrast the oxygen consumption of slices incubated in potassium-free medium with 10 mmol.1-1 ouabain was markedly reduced (by 32 +/- 6%), while such slices lost 180 +/- 15 mmol of potassium/kg dry weight. These disproportionate effects on potassium loss and inhibition of oxygen consumption suggest that in renal cortical slices the loss of potassium in low potassium medium is not primarily due to inhibition of the conventional sodium pump.     

ANOMALIES IN THE ABSORPTION SPECTRUM AND BLEACHING KINETICS OF VISUAL PURPLE

Visual purple from winter frogs shows an intermediate yellow color during bleaching by light; summer extractions do not. This seasonal effect can be duplicated by variations in the hydrogen ion concentration and in the temperature of the solutions. Increasing the pH approximates the summer condition, while decreasing the pH approximates the winter condition. Temperature has no effect on the bleaching of alkaline solutions but greatly influences acid solutions. At low temperatures the bleaching of add solutions resembles the winter condition, while at higher temperatures it resembles the summer condition. A photic decomposition product of frog retinal extractions is an acid-base indicator: it is yellow in acid and colorless in alkaline solution. Its color is not dependent upon light. The hydrogen ion concentration of visual purple solutions does not change under illumination, nor is there a difference in the pH of summer and winter extractions. Bile salt extractions of visual purple are usually slightly acid. The conflicting results of past workers regarding the appearance of "visual yellow" may be due to seasonal variation with its differences in temperature, or to the presence of base in the extractions. It is also possible that vitamin A may be a factor in the seasonal variation. The photic decomposition of visual purple in bile salts solution, extracted from summer frogs, follows the kinetics of a first order reaction. Visual purple from winter frogs does not conform to first order kinetics. Photic decomposition of alkaline, winter visual purple extractions also follows a first order equation. Acid, winter extractions appear to conform to a second order equation, but this is probably an artefact due to interference by the intermediate yellow.     

CLINICAL POTASSIUM PROBLEMS

Alterations in serum potassium are common in many diseases. In a series of 390 determinations of serum potassium, the levels found were low in 24 per cent and high in 2.6 per cent.The major causes of low serum potassium are (1) decreased potassium intake due to intravenous feedings which do not contain potassium; (2) increased loss of potassium in the urine due to accelerated tissue breakdown, or renal lesions; (3) loss from the gastrointestinal tract due to diarrhea, or fistulae, and (4) shift between serum and cells, due to metabolic causes, drugs or changes in pH.The major cause of high serum potassium is uremia with renal retention.Clinical symptoms and signs of low body potassium include muscle weakness and paralysis, which may lead to death in respiratory failure if not corrected, tachycardia, gallop rhythm, dilatation of the heart. The electrocardiogram shows inverted, low amplitude, or isoelectric T waves and a prolonged QT interval.Potassium chloride orally, subcutaneously or intravenously is recommended for use in the treatment of potassium deficits. It should not be used in the presence of oliguria or anuria or dehydration. The amounts of potassium necessary to correct deficits vary widely and cannot be predicted from the serum level. Special reference is made to the prevention and therapy of potassium deficits in diabetic acidosis.High serum potassium levels are difficult to correct. Suggested measures are administration of glucose, insulin or calcium, gastric or peritoneal lavage or use of the artificial kidney.     

氯酸钾对花生生长的毒害效应

氯酸盐是一类毒性强的氧化剂,曾被作为非选择性除草剂和脱叶剂大量施用,近年来,氯酸钾作为产期调控剂也在龙眼(Dimocarpus longan)反季节生产中大量应用。然而,氯酸根离子强氧化性对生物体有明显的毒害效应,其残留及次生污染物对水体和土壤环境也存在较强的污染效应。龙眼园通常间种花生(Arachis hypogaea),该文实验研究了氯酸钾和氯化钠对花生生长的毒害效应。结果表明,当浸种溶液的KClO₃浓度高于50 mg·L^-1时,花生种子的发芽率和胚根长度大大降低,幼芽的电解质渗漏率和过氧化氢酶活性显著升高;当土壤中KClO₃浓度高于50 mg·kg^-1时,会使花生幼苗叶片质膜透性增大,而硝酸还原酶活性、叶绿素含量和根系活力显著降低,氯酸钾的毒害效应远远超过氯化钠的盐害效应,高于50 mg·kg^-1的KClO₃还能使花生荚果期植株的光合速率、蒸腾速率、叶绿素含量、根系活力、生物量合成和根瘤菌的数量显著降低。结论是,土壤中的KClO₃浓度高于50 mg·kg^-1时,即会显著影响花生植株的正常生长。     

THE EFFECT OF POTASSIUM ON THE ACID METABOLISM OF SURVIVING SKELETAL AND CARDIAC MUSCLES OF THE FROG

The potassium contraction of skeletal muscle and relaxation of cardiac muscle have been correlated with the carbon dioxide and total acid production of these tissues. 1. The immersion of surviving sartorius muscles of the frog in isotonic potassium chloride solution causes a marked increase in the rate of acid production. 2. It is probable that carbon dioxide is the principal acid involved in the above effect. 3. The immersion of surviving cardiac muscle of the frog in isotonic potassium chloride solution causes a pronounced depression in the rate of survival acid production. 4. Reasons are given for believing that these changes in metabolism may be independent of the stimulation and inhibition of contraction which potassium simultaneously produces in these tissues.     

Release of beta-alanine from the frog skeletal muscle determined by thin-layer chromatography

The release of beta-alanine from the resting and contracting frog sartorius muscles was demonstrated by the two-dimensional thin-layer chromatography. The release of beta-alanine from indirectly stimulated muscles of frogs in winter was about 230% higher than at rest. When synaptic transmission was blocked by d-tubocurarine the release of beta-alanine from directly stimulated muscles did not exceed the release at rest. Thus, activation of neuromuscular synapse leads to increased beta-alanine release from contracting muscle.     

Affinities or Apparent Affinities of the Transport Adenosine Triphosphatase System

The interactions of potassium ions and ATP on transport ATPase activity are discussed, and the interpretation of these interactions is shown to be often ambiguous. Caldwell''s (1968) Physiological Review model is discussed with particular reference to the observed kinetics of sodium: sodium exchange in red cells. Recent experimental work on the properties of the ouabain-sensitive component of potassium efflux from red cells is described. This component of efflux occurs only if either sodium or potassium are present in the external medium, but the effects of external sodium and potassium are not additive. The relation between ouabain-sensitive potassium efflux and the external concentration of sodium (in a potassium-free medium) or of potassium (in low- and high-sodium media) are described. When starved sodium-poor red cells are poisoned with iodoacetamide, loaded with phosphate, and incubated in high-sodium potassium-free media, the ouabain-sensitive efflux of potassium appears to be accompanied by the reversal of the entire ATPase system. About two to three potassium ions leave by the ouabain-sensitive route for each molecule of ATP synthesized. If potassium is present in the external medium, no ouabain-sensitive synthesis of ATP occurs and the ouabain-sensitive efflux of potassium presumably involves the reversal of only the last part of the ATPase system.