Role of tropomyosin in the sensitivity of (Na+ + K+) ATPase to ouabain]

EDTA treatment of isolated plasma membranes from MF2S cells increased 1,000 fold the sensitivity of (Na+ + K+) ATPase activity to ouabain. The original sensitivity of the enzyme to the drug is recovered after addition of tropomyosin together with Ca++ ions to the treated membranes.     

Inhibition of hepatic Na +, K + -adenosinetriphosphatase in taurolithocholate-induced cholestasis in the rat.

Na +, K + -adenosinetriphosphatase (Na +, K + -ATPase) activity was decreased in liver plasma membranes from rats in which cholestasis had been induced by i.v. administration of sodium taurolithocholate (5 mumoles/100 g b. wt). Incubation of liver plasma membranes with taurolithocholate (10--1300 muM) caused significant and dose dependent reductions of Na +, K + -ATPase activity at taurolithocholate concentrations above 100 muM. These findings lend support to the hypothesis that cholestasis induced by monohydroxy bile acids is at least partially the result of an inhibition of hepatic Na +, K + -ATPase activity.     

Inhibition of diuresis in the tsetse fly (Glossina morsitans) by ouabain and acetazolamide.

Acetazolamide and ouabain, metabolic inhibitors which interfere with certain membrane transport systems, reduce the rate of water elimination by male Glossina morsitans morsitans. The results suggest that water is transported across membranes during diuresis and that a ouabain sensitive Na+K+ATPase and an acetazolamide-sensitive carbonic anhydrase are involved in diuresis.     

Purification from a total cellular lysate of a protein modifying the sensitivity of Na+/K+ ATPase to ouabain]

A protein (32,000 dalton) has been purified from M2FS cells derived from the murine plasmocytoma MOPC 173. Like tropomyosin, this protein when added with Ca++ to EDTA-treated plasma membranes prepared from the same cell, induced a drastic increase in the Na+/K+ atpase resistance to ouabain.     

Effects of inhibition and stimulation of Na+-K+ active transport on the resting membrane input conductance of the guinea-pig ventricle

The effects of inhibition by ouabain and stimulation by high frequency drive of the sarcolemmal Na+-K+ active transport system on the resting input conductance (gi) of guinea-pig ventricular muscles were determined. Although both pump inhibition and stimulation were associated with changes in electrophysiological properties of the muscles, neither had a significant effect on gi.     

Inhibition of microsomal Na+ K+ ATPase of the brain by extracts of Mansonia altissima]

Water extracts of the bark of Mansonia altissima var altissima inhibit the activity of the Na+, K+-ATPase of Rabbit brain microsomes. The kinetics of hydrolysis of ATP show non-competitive inhibition analogous to that produced by ouabain.     

Na-pump sites in the oviduct of Salamandra salamandra (L.) (Amphibia, Urodela)

Using 3H-ouabain autoradiography, Na+-K+-ATPase has been localized on the basolateral membranes of ciliated and nonciliated cells in the oviduct (pars recta, p. convoluta I, II, III) of the European fire salamander, Salamandra salamandra. The mucous and seromucous gland cells of the p. convoluta I, II, III, however, do not show any significant labelling. An asymmetrical distribution of ouabain binding sites is a main feature of transporting epithelia.     

Influence of medium amino acids on the ouabain sensitive and insensitive 86Rb+-fluxes in HeLa cells

Components of the 86Rb+-influx in HeLa cells were investigated in Joklik minimal essential medium, or in Earle's balanced salt solution with and without medium amino acids. The presence of amino acids led to the stimulation of the ouabain sensitive 86Rb+-uptake and inhibition of the diuretic-sensitive and residual 86Rb+-fluxes. These results show that the presence of amino acids is an important regulator of the K+/Rb+-fluxes under normal conditions in growth medium.     

Inhibition of sarcolemmal Na, K, Mg ATPase from the guinea pig heart is not compatible with a homogeneous population of non-interacting ouabain receptors

The inhibition of sarcolemmal Na, K, Mg ATPase from the guinea pig heart by ouabain was evaluated with a coupled enzyme assay. Models of negative cooperativity and of two independent receptors fitted the inhibition data equally well. The analysis was not compatible with a homogeneous population of non-interacting ouabain receptors.     

Inhibition of sarcolemmal Na,K, Mg ATPase from the guinea pig heart is not compatible with a homogeneous population of non-interacting ouabain receptors

Summary The inhibition of sarcolemmal Na, K, Mg ATPase from the guinea pig heart by ouabain was evaluated with a coupled enzyme assay. Models of negative cooperativity and of two independent receptors fitted the inhibition data equally well. The analysis was not compatible with a homogeneous population of non-interacting ouabain receptors.Acknowledgments. The technical assistance of P. Mayr and G. Ruhland is gratefully acknowledged.     

Inhibitory effect of ouabain on in vitro and in vivo gastric acid secretion in the frog and the rat

The in vitro and in vivo effects of ouabain on gastric acid secretion in the frog and the rat, the 2 species known to have different sensitivity to ouabain, were studied. It was found that ouabain was a potent inhibitor of histamine-stimulated acid secretion in the isolated frog gastric mucosa. Ouabain administered i.v. at dose levels far below the lethal range also produced a marked and significant reduction of histamine-stimulated gastric acid secretion in the anesthetized frogs and rats. It is considered that the inhibitory effect of ouabain on acid secretion could be partly related to its specific antagonizing action on the Na+ -K+ -ATPase in the gastric mucosa.     

Vascular reactivity in hypertension: Altered effect of ouabain

Summary Ouabain inhibits the relaxing effect of Ca²⁺ (but not of Mn²⁺) on contractile responses in tail artery strips isolated from spontaneously hypertensive and normotensive rats. The magnitude of ouabain inhibition was greater in vascular strips from hypertensive rats suggesting a significant difference in basic membrane function in hypertensive vascular smooth muscle.These studies were supported by National Heart, Lung, and Blood Institute grants HL-03765 and HL-18575-02-03. R.C. Webb is a Postdoctoral Research Fellow of the Michigan Heart Association.     

Comparative effects of ouabain, natriuretic factor and ammonium chloride in the toad urinary bladder

Electrical changes and direct effects on Na-K ATPase activity induced by an endogenous digitalis-like natriuretic factor (NF), NH4Cl and ouabain were studied in toad bladders. NF inhibited the SCC and the Na-K ATPase activity in a similar manner to ouabain, but induced a greater increase in calculated direct current resistance (R) (p less than 0.05). NH4Cl was a weak inhibitor of Na-K ATPase activity, although it produced steeper SCC inhibition slopes than those observed with ouabain or NF (p less than 0.01). The data suggested the same mechanism of action of NF and ouabain on the sodium pump, with an additional effect of the former on apical sodium permeability of the cells and/or closure of paracellular routes leading to an increased tissue resistance. In contrast, the effects of NH4Cl were mostly compatible with intracellular inhibition of apical sodium entry into the cell.     

Synaptosomal adenosine triphosphatase (ATPase) inhibition by organophosphates.

Chicken spinal cord adenosine triphosphatases (both Na+, K+ stimulated and ouabain insensitive) were inhibited by tri-o-tolyl phosphate (TOTP, a neurotoxic organophosphate which is not a cholinesterase inhibitor) and mevinphos (a non-neurotoxic compound but inhibitor of cholinesterases). The inhibition was concentration and time dependent, with an initial rapid drop in activity followed by a gradual exponential decline.     

Effect of ouabain on volume regulation of rabbit kidney cortex slices in hypo-osmotic media

The volume regulation process at work in rabbit kidney cortex slices submitted to hypo-osmotic media show both a swelling limitation and a volume readjustment phase. Swelling limitation is Na+ dependent and is blocked by ouabain 10(-3) M. There is, however, no need to implicate the activity of a ouabain sensitive Na+ /K+ pump in this process.     

Potentiation by taurine of the inotropic effect of ouabain and the content of intracellular Ca++ and taurine in the heart.

Under certain conditions, taurine (3.0mM) potentiated cardiac contractile response to ouabain in the normal medium. The potentiation by taurine was also observed in the low K+ medium, in which the positive inotropic effect of ouabain increased. The potentiation as seen in both media was, at least in part, due to the increase by taurine of Ca++ content in the heart. Taurine in the heart was not directly related to this potentiation.     

Inhibitory effect of taurine on decrease in the inotropic action of ouabain at high concentrations in isolated atria

Summary In vitro, taurine was shown to inhibit the decrease in the inotropic effect of ouabain at large doses in the normal and also low K⁺ medium in which this decrease in the inotropism of ouabain was facilitated. This inhibitory effect of taurine was, at least in part, due to the inhibition of the efflux of intracellular K⁺ in the isolated heart.     

The action of ouabain in promoting the release of catecholamines.

It is suggested that ouabain promotes catecholamine release by causing a rise in intracellular Na+ which, in turn, causes an elevated steady-state level of intracellular Ca2+. It is suggested that the Na+-K+-ATPase is not directly involved in exocytosis at either adrenergic or cholinergic synapses.     

Inhibition by the enterotoxin of Vibrio cholerase of meiosis reinitiation of the Xenopus laevis oocyte induced in vitro by progesterone]

Progesterone reinitiates in vitro meiosis in Xenopus laevis oocytes. This action of the hormone can be abolished by the exotoxin of vibrio cholerae. The concentration of toxin which inhibits 50% of the progesterone (10 muM) action in about 2.5 pM. Binding experiments using 125I labelled toxin demonstrated the existence of high affinity binding sites (KD approximately 0.2 nM) located probably on the surface of the oocytes.     

Effects of inhibition and stimulation of Na+−K+ active transport on the resting membrane input conductance of the guinea-pig ventricle

Summary The effects of inhibition by ouabain and stimulation by high frequency drive of the sarcolemmal Na⁺–K⁺ active transport system on the resting input conductance (g_i) of guinea-pig ventricular muscles were determined. Although both pump inhibition and stimulation were associated with changes in electrophysiological properties of the muscles, neither had a significant effect on g_i.Supported by a grant from the North Carolina Heart Association.