The role of a humoral sodium-potassium pump inhibitor in low-renin hypertension

We have recently concentrated our efforts on bioassay of plasma supernatant from animals with experimental low-renin hypertension (one-kidney, one-wrapped in dogs, and one-kidney, one-clip, and reduced renal mass in rats) for sodium-potassium pump inhibiting activity. We have observed changes compatible with inhibitory activity by using three different in vitro bioassays: 1) ouabain-sensitive 86Rb uptake by the normal rat tail artery, 2) short-circuit current in the toad bladder, and 3) membrane potential in the rat tail artery. We have also generated evidence suggesting that the humoral pump inhibitor(s) comes from or is influenced by the anteroventral third ventricle area of the brain and that it acts on the vascular smooth muscle cell at least in part by depolarizing the membrane. These findings are compatible with our 1976 hypothesis in which we proposed that in volume-expanded hypertension there is a circulating agent that suppresses cardiovascular membrane Na+,K+-ATPase, which results in reduced activity of the Na+-K+ pump and hence increased contractility of heart, arteries, and veins and that in blood vessels the increased contractility may be secondary to depolarization. We attempt to relate these findings to those in the literature on monovalent ion transport in blood cells of hypertensive subjects.     

The role of humoral agents in volume expanded hypertension.

It has long been known that increased salt intake or decreased salt excretion leads to elevated blood pressure and increased extracellular salt and volume. The blood pressure rises slowly and for this reason and others cannot be explained by the increased salt or volume per se. Rather it appears that pressure rises as a result of some indirect effect of the increased volume. Autoregulation has been implicated, but reasons are presented which make this an unlikely possibility. Recent evidence suggests that some humoral agent or agents, operating through the Na⁺?K⁺ pump in cardiovascular muscle, participate in the genesis of the volume expanded hypertensions.     

Humoral and neurogenic factors in two-kidney renovascular hypertension

A "bolus" dose (110 microgram) of the angiotesin II (A II)-blocker 1-Sar-8-Ala-A II (Saralasin, S) followed by its slow rate infusion (5 microgram/min/rat) for thirty min, was injected before and after the complete ganglionic blockade by pentolinium (P) in unanaesthetized unilaterally clipped renal hypertensive rats (the opposite kidney remained untouched). Pentolinium was also injected like a "bolus" dose (3 mg) followed by slow infusion (0.1 mg/min/rat) for thirty min. The observations were made until the fifth week after clipping the left renal artery. A consistent maximal hypotensive response was observed after the "bolus test" with both drugs. When S was the first drug injected, an inverse correlation was found between the percent decrease in arterial pressure (BP) by S and the percent decrease in BP by P (r = --0.83, P < 0.01, n = 8). Thus whenever a greater hypotensive effect was obtained by S, a smaller neural pressor component remained to be blocked by P. On the other hand, when P was the first drug injected a lesser A II pressor component remained to be blocked by S in the hypertensive rats. The results suggest that a considerable A II pressor effect in two-kidney renovascular hypertension is mediated via neurogenic mechanisms from the first week. A direct pressor vasoconstriction was found to be significant in cases with very high plasma-renin activity.     

Cell potential and the sodium-potassium pump in vascular smooth muscle.

An electrogenic sodium-potassium pump appears to contribute materially to the steady-state potential and to certain of the transient potential responses of vascular smooth muscle. Since changes in cell potential in turn can lead to changes in contractile state, the pump is implicated in some of the constriction-dilation responses of blood vessels. The vasodilator action of potassium is explainable, for instance, through an effect on cell potential if (and only if) an electrogenic pump is assumed to be extruding sodium at a faster rate than it takes up potassium. This is supported by the observation that ouabain, an inhibitor of Na,K-ATPase activity, will eliminate or reverse the vascular effect of potassium. Furthermore, when the in vivo and in vitro effects on vascular smooth muscle of altered extracellular potassium concentration are compared to calculated cell potentials based on a model that includes an electrogenic pump, the experimental findings are shown to be logical and predictable.     

Role of potassium channels, the sodium-potassium pump and the cytoskeleton in the control of dog sperm volume

Response to osmotic shock is an important aspect of mammalian sperm physiology. In this study we recorded volume changes of dog spermatozoa at 39, 33, and 25 degrees C under isotonic conditions and following hypotonic shock. Cell volume measurements were performed electronically in saline solutions of 300 and 150 mOsmol kg(-1), and Percoll-washed preparations were compared with unwashed samples. The involvement of potassium channels in volume control was tested by treatment with quinine, while the involvement of the plasma membrane Na(+)-K+ pump was tested by treatment with ouabain. The role of the cytoskeleton was investigated by treatment with colchicine and cytochalasin D. The number of cell populations observed varied with temperature and tonicity. In both types of sperm preparations, between two and three populations were present under isotonic conditions at 25 degrees C whereas at 39 and 33 degrees C only one population was detected. Hypotonic stress at the higher temperatures caused the single population to swell, whereas at 25 degrees C it resulted in a population of cells whose modal volume was similar to that of the middle isotonic sub-population. Both quinine and the cytoskeletal inhibitors markedly increased swelling both under hypotonic conditions at 39 degrees C and under isotonic conditions at 25 degrees C. However, little or no effect of ouabain was observed. We conclude that in dog spermatozoa swelling in response to hypotonic conditions is minimised through the activity of potassium channels and the presence of an intact cytoskeletal network. Under isotonic conditions at 25 degrees C, a considerable proportion of the sperm population is already swollen; this swelling varies between individual males and appears to be due to lowered cytoskeletal and potassium channel activity.     

On control of the function of the sodium-potassium pump in malignant cells

This paper introduces a double-layer enzyme-membrane model representing the Na⁺−K⁺ pump in living cells. We present a mathematical solution to the problem of controlling the sodium flux in malignant cells, where an inhibitor exists in the outer layer of the membrane. We give an algorithm for the numerical resolution of this problem of optimal control with illustrations. Finally, we point out the biological importance of this study.     

Effect of palytoxin on the sodium-potassium pump: model and simulation

We propose a reaction model for the palytoxin-sodium-potassium (PTX-Na(+)/K(+)) pump complex. The model, which is similar to the Albers-Post model for Na(+)/K(+)-ATPase, is used to elucidate the effect of PTX on Na(+)/K(+)-ATPase during the enzyme interactions with Na(+) and/or K(+) ions. Conformational substates and reactions for the pump are incorporated into the Albers-Post model to represent enzymes with or without bound PTX. A mathematical model based on the reaction scheme is used in simulations modeling experimental studies of PTX-induced ionic currents. Our simulations suggest that (i) extracellular Na(+) as well as K(+) promotes PTX-induced channel blockage; (ii) extracellular K(+) accelerates PTX unbinding; and (iii) K(+) occlusion in the PTX-pump complex is essential for describing the PTX-induced current dynamics.     

Decreased sodium-potassium pump activity in isolated hypertrophied feline ventricular myocytes

The activity of the Na-K pump was assessed in normal and hypertrophied isolated feline myocytes by measuring ouabain-sensitive 42-K uptake. Right ventricular hypertrophy was produced in feline myocardium by placing a constricting band around the pulmonary artery of adult cats. High yields of calcium tolerant myocytes were isolated from the right and left ventricle of banded and sham operated animals. Intracellular sodium (Na) and potassium (K) concentrations (mM) were not significantly different (P greater than 0.5) in normal (Na: 13.2; K: 133.4) and hypertrophied (Na: 12.3; K: 127.5) myocytes. Morphometric analysis demonstrated a 26% increase in width and a 42% increase in volume of hypertrophied myocytes, however, the sarcomere length (1.9 mu) was not different in both cell types. The rate constant (k, min-1) describing 42-K uptake and the calculated total K influx (I, pmol/cm2/sec) were not significantly different (P greater than 0.5) in normal (k = 0.059; I = 15.9) and hypertrophied (k = 0.062; I = 15.3) cells. Ouabain-sensitive (active) K influx, a measure of Na-K pump activity, was maximally inhibited at 10(-4)M ouabain in both cell types. At this concentration, ouabain-sensitive K uptake was decreased 23.5% in hypertrophied myocytes compared to control. The decrease in active K influx may be due to a decrease in the activity of the Na-K ATPase and/or to a reduction in the passive movement of sodium and potassium down their electrochemical gradients.     

The effect of hyperthermia on the sodium-potassium pump in Chinese hamster ovary cells

The effect of hyperthermia on the Na+-K+ pump was determined by measuring influx and efflux of 86Rb+ in Chinese hamster ovary cells from 31 to 50 degrees C. The maximum initial rate of ouabain-sensitive influx increased with temperature between 31 and 45 degrees C although Km increased significantly above 37 degrees C, implying a diminished affinity of the transport protein for its substrate. The changes in the kinetics of influx at temperatures up to 45 degrees C were rapidly reversible on return to 37 degrees C. Above 45 degrees C an irreversible decrease in 86Rb+ uptake was observed. Efflux of 86Rb+ increased from 31 to 40 degrees C but above 43 degrees C showed a small but significant decrease. The study of 86Rb+ influx after varying times of exposure to elevated temperatures showed that the Na+-K+ pump remains functional in cells which are reproductively dead. We have shown that although the kinetics of K+ transport are sensitive to temperature changes in the range used in clinical hyperthermia, the inactivation of the Na+-K+ pump is not a primary event in cell killing.     

Humoral vasopressor agents in primary hypertension

In essential hypertension and in the spontaneously hypertensive rat, there are indications for a yet unidentified vasopressor agent which may play an important role in pathogenesis. It has been demonstrated in the spontaneously hypertensive rat by parabiosis, cross-circulation, cross-transplantation of kidneys and by injection of hypertensive plasma fractions. In plasma from essential hypertensives, an agent sensitizing the vasculature to pressor hormones and a substance with direct vasopressor actions have been found. The relationship of the hypertensive factor to the natriuretic hormone is not clear.     

Arachidonate has a positive effect on the electrogenic sodium-potassium pump in the mouse diaphragm

Sodium arachidonate 5 X 10(-5) mol X l-1 shortened the time course of hyperpolarization caused by the electrogenic Na+-K+ pump in intact muscle fibres in the mouse diaphragm preincubated in a K+-free physiological solution. Contrary to experiments on membrane fragments, no inhibition of the ouabain-sensitive Na+-K+ ATPase was observed. It is unlikely that the arachidonate may be identical with the endogenous "ouabain-like" substance (Bidard et al. 1984).     

Regulation of the sodium-potassium pump in cultured rat skeletal myotubes by intracellular sodium ions

The properties of the Na-K pump and some of the factors controlling its amount and function were studied in rat myotubes in culture. The number of Na-K pump sites was quantified by measuring the amount of [3H]ouabain bound to whole-cell preparations. Activity of the pump was determined by measurement of ouabain-sensitive 86Rb-uptake and component of membrane potential. Chronic treatment of myotubes with tetrodotoxin (TTX), which lowers [Na]i, decreased the number of Na-K pumps, the ouabain-sensitive 86Rb uptake, and the size of the electrogenic pump component of Em. In contrast, chronic treatment with either ouabain or veratridine, which increases [Na+]i, resulted in an elevated level of Na-K pump sites. This effect was blocked by inhibitors of protein synthesis. Neither rates of degradation nor affinity of pump sites in cells treated with TTX, veratridine, or ouabain differred from those in control cells. The number and activity of Na-K pump sites were unaffected by chronic elevation in [Ca]i or chronic depolarization. We conclude that alterations in the level in intracellular Na ions play the major role in regulation of Na-K pump synthesis in cultured mammalian skeletal muscle.