Effect of free fatty acids and cholesterol in vitro on liver plasma membrane-bound enzymes

The effect of cholesterol and fatty acid treatment in vitro was tested on rat liver plasma membrane-bound enzymes and lipid fluidity. The observed alterations of membrane fluidity affect both (Na+-K+)-ATPase and Mg2+-ATPase activities but not 5'-nucleotidase; basal adenylate cyclase as well as its hormonal sensitivity were differentially affected by changes of membrane microenvironment.     

Characterization of the (Na+-K+)-ATPase from endometrial plasma membranes of immature mammals]

The (Na+-K+)-ATPase in plasma membrane from Mammiferous endometrium is characterized by the Mg/ATP ratio equal to one, and by a distinct affinity for Na+ (1.3 mM) and K+ (2 mM). The activity is maximum for pH 7.4-7.5 in presence of Mg++ 2mM and ATP 2 mM, Na+ 140 mM and K+ 10 mM.     

Alteration of erythrocyte (see article) -ATPase by replacement of cholesterol by desmosterol in the membrane.

Cholesterol of red blood cells (RBC) is readily exchanged by desmosterol and vice versa. The resulting alteration in the sterol composition influences the specific (Na+ plus K+)-ATPase activity. It is suggested that this effect is due to an altered membrane fluidity.     

Solubilization and characterization of a ouabain-sensitive protein from transverse tubule membrane-junctional sarcoplasmic reticulum complexes (TTM-JSR) in cat cardiac muscle

A new glycoprotein of 31,500 dalton, which has a high affinity for ouabain, and is independent of (Na+-K+)-ATPase, was solubilized from transverse tubule membrane and junctional sarcoplasmic reticulum complexes (TTM-JSR) of cat cardiac muscle. This protein could be extracted only in small amounts from sarcolemma-plasma membrane (SLM-PL) fragments, suggesting that it indeed originates from the TTM-JSR.     

The (Na+ + K+)-ATPase activity in brain of Quaking mice.

The (Na+ 4 K+)- and Mg2+-dependent ATPase distribution in several brain areas has been investigated in Quaking mutant mice characterized by myelin deficiency. A marked decrease of (Na+ + K+)-ATPase activity has been found in limbic structures, hypothalamus and cerebellum. The Mg2+-dependent activity did not change. A possible involvement of the impairment of the (Na+ + K+)-ATPase activity in the seizure susceptibility of this mice is discussed.     

The effect of physostigmine on (Na+ + K+)-ATPase activity in different rat brain regions

The activity of (Na+ + K+)-ATPase and acetylcholine esterase were followed in rat brain cerebral cortex, caudate, thalamus, hippocampus and medulla after i.v. administration of physostigmine. Both enzymes were found to be inhibited in a dose-dependent manner. The most pronounced inhibition of (Na+ + K+)-ATPase was found in caudate, where the highest activity of acetylcholine esterase is found.     

Insulin effect in vitro on human erythrocyte plasma membrane

Summary The effect of porcine insulin has been tested in vitro on human erythrocyte plasma membrane (Na⁺–K⁺) and Mg²⁺-ATPase activities as well as on membrane fluidity. The results indicate that the hormonal treatment significantly inhibits (Na⁺–K⁺)-ATPase activity, and at the same time decreases membrane fluidity.This investigation has been supported by Consiglio Nazionale delle Richerche, Rome, Italy.     

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.     

Cytochemical localization of the K+ regulation interface between blood and brain.

Phosphatase activity identified with Na+-K+-ATPase was localized at the basal surface of cerebral cortical capillary endothelium by perfusion with a p-nitrophenyl phosphate-strontium medium. The relationship of this to the blood-brain barrier to K+ is discussed.     

Imbalance in the activities of alkaline phosphatase and Na+-K+-ATPase in the brain of experimentally induced phenylketonuric squirrels (Funambulus palmarum).

Phenylketonuric squirrels have shown marked inhibition of alkaline phosphatase in the olfactory lobes and cerebral hemispheres, whereas the Na+-K+-ATPase remained less altered. In the pathogenesis of phenylketonuria inhibition of alkaline phosphatase at the level of "Blood-Brain Barrier" (BBB), leads transport system to impaired functioning.     

Effect of free fatty acids and cholesterol in vitro on liver plasma membrane-bound enzymes

Summary The effect of cholesterol and fatty acid treatment in vitro was tested on rat liver plasma membrane-bound enzymes and lipid fluidity. The observed alterations of membrane fluidity affect both (Na⁺–K⁺)-ATPase and Mg²⁺-ATPase activities but not 5-nucleotidase; basal adenylate cyclase as well as its hormonal sensitivity were differentially affected by changes of membrane microenvironment.This investigation was partially supported by the Italian National Research Council.     

Solubilization and purification of Na+-K+ ATPase of human erythrocyte mambranes using sodium deoxycholate]

The (Na+-K+)-dependant ATPase prepared from human erythrocyte ghosts has been purified with sodium deoxycholate in a high ionic medium. After the purification, 60% of the native activity bound to the membranes was recovered.     

Decreased monosaccharide transport in renal brush-border membrane vesicles of spontaneously hypertensive rats

Na(+)-dependent D-glucose and D-galactose transport were studied in brush-border membrane vesicles (BBMVs) from kidney cortex isolated from both spontaneously hypertensive rats (SHR) and their normotensive genetic control Wistar-Kyoto (WKY) rats. Initial rates and accumulation ratios of Na(+)-dependent D-glucose and D-galactose transport were significantly lower in SHR compared with WKY, the observed decreases being similar for both substrates. To explain the reduction in sugar transport by renal BBMVs, the density of Na(+)-dependent sugar cotransporters was studied in BBMVs from kidney cortex isolated from SHR and WKY rats. Phlorizin-specific binding and Western blot analysis indicated a reduction in the density of the cotransporters in SHR relative to WKY rats. This reduction was similar to those found for the initial rates and accumulation ratios for D-glucose and D-galactose in SHR. Na+ uptake, studied using 22Na+, was significantly increased in SHR, so the observed reduction in sugar transport could be due to disruption of the Na+ gradient between renal BBMVs in SHR. Furthermore, a significant decrease in the activity of Na(+)-K(+)-ATPase was observed in SHR. In conclusion, changes in the density of the Na(+)-dependent sugar cotransporter and in the Na+ gradient across the brush-border membranes might be involved in the observed reduction in sugar transport by renal BBMVs from SHR.     

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.     

Possible role of intestinal alkaline phosphatase activity in thiamine transport.

Thiamine deficiency caused a marked decrease of intestinal alkaline phosphatase (al-Pase) activity, but had no effect on the Ca++-ATPase activity and Ca++-absorption in rats. The al-Pase activity was significantly decreased 1 h after oral administration of ethanol at 0.5 and 2.5 g/kg. In contrast, Mg++-, Ca++-and (Na+ + K+)-ATPase activities did not change after the administration of ethanol. These findings show that the al-Pase activity, unlike the Ca++-ATPase activity, is not related to Ca++-absorption. A possible role of al-Pase activity in the active transport of thiamine in the intestine was discussed.     

Phospholipid composition of cardiac (Na+ + K+)-ATPases from various species

There is a difference in phospholipid composition of cardiac (Na+ + K+)-ATPase preparations between species which are sensitive to ouabain and those which are not. Sphingomyelin is higher and phosphatidylcholine is lower in the enzymes from sensitive species than in those from insensitive ones. Lysophosphatidylcholine is detectable only in the latter preparations.     

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.     

Effects of EDTA on (Na+-K+) ATPase bound to human erythrocyte membranes]

Membranes preincubation with high EDTA concentration induced an alteration on (Na"-K+) ATPase kinetic behaviour. This effect can be related to a change in calcium content tightly bound to the membranes.     

A histochemical demonstration of the Na+ + K+-ATPase activity in the thyroid and the effect of cyclic adenosine monophosphate (c-AMP).

With a suitable modification of the Farquhar and Palade technique the Na+ + K+-ATPase activity in guinea-pig thyroid is demonstrated. The addition of c-AMP (5 X 10(-6) M or 1.5 X 10(-5) M) to the incubation media produced an apparent intensification of the Na+ + K+ -ATPase activity in the thyroid.     

Effect of mycotoxin (T-2 toxin) on catecholamine and Na+, K+-ATPase levels in rat epididymis

The effect of mycotoxin (T-2 toxin) on catecholamines and Na+, K+-ATPase activities in rat epididymis has been evaluated. Dopamine and norepinephrine levels were significantly elevated in the caput and corpus regions whereas their levels remained unchanged in the caudal part of the epididymis. Na+, K+-ATPase activity was significantly increased in all the three regions of rat epididymis as a result of the toxin treatment. These changes may suggest an adverse effect on epididymal functions in rats.