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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We hypothesized that viral mediated transfer of Na+,K+-ATPase subunit genes to alveolar epithelial cells to overexpress Na+, K+-ATPase could increase Na+,K+-ATPase function. We produced replication-deficient human type 5 adenoviruses that contained cytomegalovirus (CMV)-driven cDNAs for the rat alpha1 and beta1 subunits of Na+,K+-ATPase (AdMRCMValpha1 and AdMRCMVbeta1, respectively). These viruses were used to transduce human adenocarcinoma cells (A549) in culture. Na+,K+-ATPase function was increased by 2.5-fold in the AdMRCMValpha1-infected cells. Sham and AdMRCMVbeta1-infected cells, and cells infected by a CMV-driven beta-galactosidase-expressing adenovirus, had no increases in Na+, K+-ATPase activity. A549 cells infected with multiplicities of infection of 10-200 of AdMRCMValpha1 demonstrated expression of a rat alpha1 mRNA and increased alpha1 protein; no change in beta1 message or protein was noted. Ouabain sensitivity was measured in A549 cells following infection with AdMRCMValpha1. In contrast to controls, AdMRCMValpha1-infected cells demonstrated two IC50s. The first was similar to the IC50s of the controls; the second IC50 was 2 logs greater than the first, consistent with the presence of both the rat and human alpha1 isozymes. These results demonstrate for the first time that adenoviruses can be used to augment Na+,K+-ATPase function.
Am J Respir Cell Mol Biol 1998 Jun
PMID:Overexpression of the Na+,K+-ATPase alpha1 subunit increases Na+,K+-ATPase function in A549 cells. 961 78

The changes of Na,K-ATPase activity and its regulation have been investigated in the renal cortex and its basolateral membrane of aldosterone-induced hypertensive rat. Ouabain-sensitive Na,K-ATPase activity and [3H]ouabain-binding site (Bmax) in the hypertensive rat were significantly increased than those in the control. The levels of Na,K-ATPase alpha 1- and beta 1-subunit mRNA of the renal cortex in hypertensive rat were more increased than those in the control, and their increases were repressed by actinomycin-D. These results suggest that the increase of Na,K-ATPase activities and ouabain binding sites in aldosterone-induced hypertensive rat may be correlated with transcriptional regulation of Na,K-ATPase gene expression.
Biochem Mol Biol Int 1998 Aug
PMID:Aldosterone stimulates Na,K-ATPase activity in basolateral membrane of rat kidney. 973 52

The changes of Na,K-ATPase activity and its regulation have been investigated in the renal cortex of 1-clip-1-kidney hypertensive rat. Ouabain-sensitive Na,K-ATPase activity (Emax) and [3H]ouabain-binding site (Bmax) in the hypertensive rat were slightly increased than those in the control. The levels of Na,K-ATPase alpha 1- and beta 1-subunit mRNA of the renal cortex in hypertensive rat were more increased than those in the control. Their increases were repressed by actinomycin-D, but not altered or more increased by cycloheximide. These results suggest that the increase of Na,K-ATPase activities and ouabain binding sites in 1-clip-1-kidney hypertensive rat may be correlated with the increases of gene expression in transcription level and/or of mRNA stability of Na,K-ATPase.
Biochem Mol Biol Int 1998 Nov
PMID:Regulation of Na,K-ATPase activity in renal basolateral membrane of 1-clip-1-kidney hypertensive rat. 984 26

Ascorbic acid serves a vital role as a pre-eminent antioxidant. In animals, it has been shown to be concentrated in granulosa and theca cells of the follicle, in luteal cells of the corpus luteum, and in the peripheral cytoplasm of the oocyte. We have previously identified hormonally-regulated ascorbic acid transporters in rat granulosa and luteal cells, and herein present preliminary evidence for the presence of a transporter for ascorbic acid in human granulosa-lutein cells. Granulosa-lutein cells were obtained from the follicular fluid of patients undergoing in-vitro fertilization. Following an overnight incubation, the cells were incubated with [14C]-ascorbic acid (0.15 microCi; 150 microM) and ascorbic acid uptake was determined. The uptake of ascorbic acid was saturable with a Michaeli's constant (Km) and maximum velocity (Vmax) of 21 microM and 3 pmol/10(6) cells/min respectively. Ouabain, low Na+ medium, and dinitrophenol significantly inhibited ascorbic acid uptake (P<0.05). Neither the presence of insulin, human chorionic gonadotrophin (HCG), insulin-like growth factor (IGF)-I, nor IGF-II affected the uptake of ascorbic acid in a statistically significant fashion. Following saturation of cellular uptake, the ascorbic acid level was estimated to be 1.04 pmoles/10(6) cells or approximately 1 mM, a high concentration similar to that seen in rat luteal cells. Active ascorbic acid transport in human granulosa-lutein cells appears to occur via a Na+ - and energy-dependent transporter, with high levels of ascorbic acid being accumulated in these cells.
Mol Hum Reprod 1999 Apr
PMID:Identification and characterization of an ascorbic acid transporter in human granulosa-lutein cells. 1032

Embryo metabolism was evaluated during re-expansion of in vitro produced bovine blastocysts collapsed with cytochalasin D (CCD) and incubated in the presence and absence of ouabain, a specific inhibitor of the Na+, K+ pump. Day 8 expanded blastocysts were treated for 2 to 4 hr with 20 microg/ml CCD. Four conditions were tested: untreated embryos and embryos collapsed with CCD and allowed to re-expand for 4 hr in the presence of 0 M, 1 nM, or 1 microM ouabain. Incubation of collapsed embryos for 4 hr in the presence of 1 nM or 1 microM ouabain significantly inhibited blastocyst re-expansion. Glucose, pyruvate, and amino lactate uptake/release were not significantly affected by ouabain treatment and did not correlate with the degree of blastocyst re-expansion. Few variations in the uptake/release of amino acids by the embryos were observed. Ouabain treatment significantly decreased oxygen uptake which directly correlated with the degree of blastocyst re-expansion. For embryos allowed to re-expand in the presence or absence of ouabain, a direct correlation was observed between the uptake of oxygen and of glucose. One mM cyanide or 2,4 dinitrophenol inhibited blastocyst re-expansion although 0.01 and 0.1 mM were ineffective. This study indicates a role for oxidative metabolism in providing the energy necessary for blastocoel expansion in the bovine. Nevertheless, blastocyst expansion is relatively insensitive to inhibition of oxidative phosphorylation indicating the ability of the bovine blastocyst to adapt to hypoxic conditions.
Mol Reprod Dev 1999 Jun
PMID:Embryo metabolism during the expansion of the bovine blastocyst. 1033 55

The interdependence between motility, respiration, ATP production, and utilization was investigated in intact spermatozoa of turbot (Psetta maxima), a marine teleost. When spermatozoa were diluted in a hyperosmotic medium (>300 mOsmol/kg), they immediately became motile, and the intracellular concentration of ATP as well as the adenylate energy charge ratio dropped concomitant with the straight-line velocity. The ADP and AMP levels increased from 1.4 to 8.0 nmole/10(8) cells and from 0.6 to 6.0 nmole/10(8) cells, respectively. Moreover, 31P-NMR spectra recorded prior to the swimming phase revealed the presence of phosphomonoesters (PMEs) and phosphodiesters (PDEs), intracellular inorganic phosphate (Pi), and phosphocreatine (PCr). At the end of the motility period, PCr, PDE, and PME decreased, while the Pi level increased markedly. Following initiation of motility, O2 consumption of spermatozoa increased from 34.9 to 124.8 O2 nmole/10(9) spermatozoa/min. FCCP, an uncoupler of oxidative phosphorylation, did not significantly affect the respiratory rate of motile spermatozoa. Ouabain, a specific inhibitor of (Na+/K+)/ATPase, slightly decreased the respiration rate of motile spermatozoa, indicating that the major part of ATP catabolism was linked to dynein ATPase. Inhibitors of the respiratory chain (KCN, NaN3, NaHCO3-, oligomycin) reduced sperm respiration, percentage of motile cells, velocity, and adenylate contents. Following the reactivation of motility of demembranated spermatozoa, KCN, NaN3, NaHCO3- altered the flagellar beat frequency, demonstrating that these respiratory inhibitors possess action sites other than mitochondria. Mitochondrial oxidative phosphorylation is highly requested to produce energy required during motion. Nevertheless it is insufficient to maintain endogenous ATP stores. A second phase of motility was induced by a transfer of exhausted spermatozoa into an ionic medium of low osmolality (200 mOsmol/kg) for 30 min. Spermatozoa, once reactivated in AM, recovered 55% of initial motility and 31% of initial fertilization rate. In hypo-osmotic medium, mitochondrial oxidative phosphorylation also induced ATP regeneration. Following activation of movement, several morphological changes were observed in the mitochondria and the midpiece.
Mol Reprod Dev 1999 Jun
PMID:Nucleotide content, oxidative phosphorylation, morphology, and fertilizing capacity of turbot (Psetta maxima) spermatozoa during the motility period. 1033 61

The antiemetic effect of granisetron, a selective 5-HT3 receptor antagonist, on ouabain-induced emesis was studied using ferrets. In order to clarify the relationship between ouabain-induced emesis and serotonin (5-HT), we examined its effects on 5-HT release from the isolated ileum. Afferent vagal nerve activity was also determined. An intravenous bolus injection of ouabain (0.1-1.0 mg/kg) produced emesis in a dose-dependent manner. Ouabain-induced emesis was inhibited by pretreatment with granisetron. In the isolated ileum, ouabain induced a concentration-dependent increase of 5-HT. This release of 5-HT was suppressed by granisetron. Increases in vagal nerve discharges were observed immediately after the intravenous administration of ouabain (0.1-1.0 mg/kg). These increases were suppressed by granisetron. Taken together, ouabain activates 5-HT release from the mucosa in the gastrointestinal tract. Released 5-HT may activate the vagal afferent nerves, resulting in vomiting. Granisetron inhibited the ouabain-induced elevation of 5-HT and vagal nerve activity. Ouabain may induce emesis as well as negative chronotropic effects by activating the vagus. Our results suggest that ouabain-induced emesis is in part mediated by the 5-HT3 receptors of the peripheral gastrointestinal tract.
Res Commun Mol Pathol Pharmacol 1998 Dec
PMID:Effects of granisetron, a selective 5-HT3 receptor antagonist, on ouabain-induced emesis in ferrets. 1034 10

Earlier studies have demonstrated that palmitoyl carnitine (PC), a long chain acyl carnitine, accumulates in the ischemic myocardium. Although perfusion of hearts with PC is known to induce contractile dysfunction which resembles ischemic contracture, the mechanisms underlying this derangement are not clear. In this study, we examined the effect of exogenous PC on the intracellular concentration of calcium ([Ca(2+)](i)) in freshly isolated cardiomyocytes from adult rat hearts. The results showed that PC elevated [Ca(2+)](i)in a dose-dependent (5-20 microm) manner; 15 microm PC evoked a marked and reversible increase in [Ca(2+)](i)without having any significant action on cell viability. The PC (15 microm)-induced increase in [Ca(2+)](i)was slightly depressed but delayed in the absence of extracellular Ca(2+). Pre-incubation of cardiomyocytes with sarcolemmal (SL) l -type Ca(2+)-channel blockers, verapamil or diltiazem, and inhibitors of SL Na(+)-Ca(2+)exchanger such as Ni(2+)or amiloride, depressed the PC-evoked increase in [Ca(2+)](i)significantly. Ouabain, a Na(+)-K(+)ATPase inhibitor, and low concentrations of extracellular Na(+)enhanced the PC-induced increase in [Ca(2+)](i). Depletion of the sarcoplasmic reticulum (SR) Ca(2+)stores by low micromolar concentrations of ryanodine (a SR Ca(2+)-release channel activator) or by thapsigargin (a SR Ca(2+)-pump ATPase inhibitor) depressed the PC-mediated increase in [Ca(2+)](i). Combined blockade of the l -type Ca(2+)channel, Na(+)-Ca(2+)exchanger and the SR Ca(2+)-pump had an additive inhibitory effect on the PC response. These observations suggest that the PC-induced increase in [Ca(2+)](i)is dependent on both Ca(2+)-influx from the extracellular space and Ca(2+)-release from the SR stores. Thus, the accumulation of PC in the myocardium may be partly responsible for the occurrence of intracellular Ca(2+)overload in ischemic heart.
J Mol Cell Cardiol 1999 Jul
PMID:Palmitoyl carnitine increases intracellular calcium in adult rat cardiomyocytes. 1040 53

An ATP-dependent transport system is responsible for the cellular extrusion of cGMP. The objective of the present study was to determine the effect of Mg2+, ATP and other nucleotides (2'-dATP, GTP and ADP), exogenous ATPase modulators (such as metavanadate, ouabain, EGTA, NEM, bafilomycin A1 and oligomycin A) on the cGMP transport. The uptake of [3H]-cGMP (1 microM) at 37 degrees C was studied in inside-out vesicles from human erythrocytes. Magnesium caused a maximal activation between 5 and 10 mM and the optimal ATP concentration was 1.25 mM with K50-values of 0.3-0.5 mM. Among other nucleotides tested, 2'-dATP (K50 of 0.7 mM) was nearly as effective as ATP, whereas cGMP accumulated slowly in the presence of GTP. ADP and metavanadate (P-type ATPase inhibitor) showed to be competitive inhibitors with Ki values of 0.15 mM and 10 microns, respectively. NEM (a sulphydryl agent) reduced the ATP-dependent uptake in a concentration-dependent manner with a Ki value of 10 microM. Ouabain (Na+/K(+)-ATPase inhibitor) had no effect. Bafilomycin A1 (V-type ATPase inhibitor) and oligomycin (F-type ATPase inhibitor) were the most potent inhibitors with Ki values of 0.7 and 1.8 microM, respectively. The present study suggests that the cellular cGMP extrusion is energized by an ATPase with a unique inhibitor profile, which clearly differentiates it from the other major classes of membrane-bound ATPases.
Mol Membr Biol
PMID:The effect of Mg2+, nucleotides and ATPase inhibitors on the uptake of [3H]-cGMP to inside-out vesicles from human erythrocytes. 1041 83

Nitric oxide generated by cardiac myocytes or delivered by drugs has been shown to regulate cardiac contractile function and has been implicated in suppressing some cardiac arrhythmias, although this remains controversial. We examined the ability of the soluble cardiac glycoside, ouabain, to trigger arrhythmic contractions in ventricular myocytes isolated from mice lacking a functional endothelial nitric oxide synthase gene (eNOS(null)). Arrhythmic activity, defined as aftercontractions, was induced with ouabain (50 micromol/L) and recorded using a video-motion detector in isolated, electrically driven single ventricular myocytes from adult eNOS(null)or from their wild-type (WT) littermates. The rate of ouabain-induced arrhythmic contractions was significantly higher in eNOS(null)myocytes than in WT myocytes. Application of the NO donor S-nitroso-acetylcysteine (SNAC) significantly diminished the frequency of arrhythmic contractions in eNOS(null)myocytes. The antiarrhythmic effect of NO, whether generated by eNOS in WT cells or by SNAC, could be partially reversed by 1H-[1,2,4]oxadiazolo-[4, 3-a]- quinoxalin-1-one (ODQ), a specific soluble guanylyl cyclase inhibitor. Ouabain significantly increased intracellular cGMP in WT but not eNOS(null)hearts, and this cGMP response was blocked by ODQ. Since cardiac glycoside- induced aftercontractions are activated by the transient inward current (I(ti)), the role of NO in ouabain (100 micromol/L)- induced I(ti)was examined using the nystatin-perforated patch-clamp technique. The frequency of ouabain-induced I(ti)was significantly higher in eNOS(null)myocytes than in WT myocytes, and this could be suppressed by SNAC. These data demonstrate that NO derived from myocyte eNOS activation suppresses ouabain-induced arrhythmic contractions by a mechanism that might involve activation of guanylyl cyclase and elevation of cGMP.
J Mol Cell Cardiol 2000 Jul
PMID:Increased susceptibility to development of triggered activity in myocytes from mice with targeted disruption of endothelial nitric oxide synthase. 1086 Jul 66


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