共查询到20条相似文献,搜索用时 15 毫秒
1.
(1) Unidirectional K+ (86Rb) influx and efflux were measured in subconfluent layers of MDCK renal epithelial cells and HeLa carcinoma cells. (2) In both MDCK and HeLa cells, the furosemide-inhibitable and chloride-dependent component of K+ influx/efflux was stimulated 2-fold by a 30 min incubation in 1 · 10?3 M ouabain. (3) Measurements of net K+ loss and Na+ gain in ouabain-treated cells at 1 h failed to show any diuretic sensitive component, confirming the exchange character of the diuretic-sensitive fluxes. (4) Prolonged incubations for 2.5 h in ouabain revealed a furosemide- and anion-dependent K+ (Cl?) outward net flux uncoupled from net Na+ movement. Net K+ (Cl?) outward flux was half-maximally inhibited by 2 μM furosemide. (5) After 2.5 h ouabain treatment, the anion and cation dependence of the diuretic-sensitive K+ influx/efflux were essentially unchanged when compared to untreated controls. 相似文献
2.
Proteoliposomes reconstituted from purified Na+ pumps show neither Ca2+ activation nor bumetanide inhibition of Rb+ uptake, suggesting that the Na+ pump does not mediate these passive fluxes. 相似文献
3.
K+ transport in 'tight' epithelial monolayers of MDCK cells. Evidence for a calcium-activated K+ channel 总被引:2,自引:0,他引:2
Measurements of 86Rb efflux across the apical and basal-lateral aspects of intact monolayers of 'high-resistance' MDCK cells mounted in Ussing chambers have been made. A transient increase in 86Rb efflux across both epithelial borders upon stimulation with adrenalineeeeeee or ionophore A23187 is observed. The increased 86Rb across the basal cell aspects is of greatest quantitative importance. Measurements of total cellular K+ contents by flame photometry of tissue extracts indicate a net loss of K+ following adrenalin addition. The effects of adrenalin and ionophore A23187 upon 86Rb efflux are abolished in 'Ca2+ -free' media. The properties of the Ca2+ -dependent increase in 86Rb efflux show similarities to Ca2+ -activated K+ conductances in other tissues, notably human red cells, including inhibition by quinine (1 mM), tetraethylammonium (25 mM) and insensitivity to bee venom toxin (apamin) (25 nM). Adrenalin is only effective when applied to the basal bathing solution suggesting that the receptors mediating adrenalin action are located upon the basal-lateral membranes. Half maximal stimulation of 86Rb efflux by adrenalin is observed at 9.1 X 10(-7) M. The action of various adrenergic receptor agonists and antagonists are consistent with adrenalin action being mediated by an alpha-adrenergic receptor. 相似文献
4.
Mouse 3T3 fibroblasts have a loop diuretic sensitive Na+ transport system, responsible for more than 50% of the total Na+ influx. This transport system is dependent on the simultaneous presence of all three ions; Na+, K+, (Rb+) and Cl- in the extracellular medium. The same requirement for these three ions was also found for the loop diuretic-sensitive K+ efflux. In addition, the sensitivities of Na+ influx and Rb+ efflux for the two loop diuretics, furosemide and bumetanide were found to be similar. The similar ionic requirement and sensitivity towards loop diuretics of the two fluxes, support the hypothesis, that this loop diuretic-sensitive Na+ influx in mouse 3T3 cells, is accompanied by the net loop diuretic-sensitive K+ efflux. 相似文献
5.
6.
Measurements of 86Rb efflux across the apical and basal-lateral aspects of intact monolayers of ‘high-resistance’ MDCK cells mounted in Ussing chambers have been made. A transient increase in 86Rb efflux across both epithelial borders upon stimulation with adrenalin or ionophore A23187 is observed. The increased 86Rb across the basal cell aspects is of greatest quantitative importance. Measurements of total cellular K+ contents by flame photometry of tissue extracts indicate a net loss of K+ following adrenalin addition. The effects of adrenalin and ionophore A23187 upon 86Rb efflux are abolished in ‘Ca2+-free’ media. The properties of the Ca2+ -dependent increase in 86Rb efflux show similarities to Ca2+-activated K+ conductances in other tissues, notably human red cells, including inhibition by quinine (1 mM), tetraethylammonium (25 mM) and insensitivity to bee venom toxin (apamin) (25 nM). Adrenalin is only effective when applied to the basal bathing solution suggesting that the receptors mediating adrenalin action are located upon the basal-lateral membranes. Half maximal stimulation of 86Rb efflux by adrenalin is observed at 9.1·10?7 M. The action of various adrenergic receptor agonists and antagonists are consistent with adrenalin action being mediated by an α-adrenergic receptor. 相似文献
7.
8.
Red cells from 3 genetically low K+ lambs exhibited: high cell K+ and K+ pump activity and 135 pumps/cell on day 3 after birth; increased cellular Na+ and a decrease of the number of pumps relative to K+ pump flux on day 17; and low cell K+, low K+ pump flux and about 40–50 pumps/cell on day 40 or later. 相似文献
9.
10.
Serum-induced net K+ influx performed by the diuretic-sensitive transport system in quiescent NIH 3T3 mouse fibroblasts 总被引:3,自引:0,他引:3
R Panet 《Biochimica et biophysica acta》1985,813(1):141-144
In serum deprived NIH 3T3 mouse cells the diuretic-sensitive transport system performs K+ self-exchange. The addition of serum which stimulates cell proliferation induces a net influx of K+, carried out by the diuretic-sensitive transport system. Thus, serum growth factors appear to induce a change in the mechanism of action of the diuretic-sensitive transporter from K+ self-exchange to an uphill transport pumping K+ into the cell. I propose here that this uphill uptake of K+ contributes to the increase of intracellular K+ content, found in the early G1 phase of the cell cycle. 相似文献
11.
Yuliya Kucherenko Joseph Browning Amanda Tattersall J Clive Ellory John S Gibson 《Cellular physiology and biochemistry》2005,15(6):271-280
Peroxynitrite is generated in vivo by the reaction between nitric oxide, from endothelial and other cells, and the superoxide anion. It is therefore pertinent to examine its effects on the membrane permeability of red blood cells. Treatment of human red blood cells with peroxynitrite (nominally 1 mM) markedly stimulated passive K+ permeability. The main effect was on a Cl(-)-independent K+ pathway, which remains unidentified. Although K+-Cl- cotransport (KCC) was stimulated, this was dependent on saline composition, being inhibited by physiological levels of glucose (IC50 4 mM), and also by sucrose and MOPS. Effects on the Cl(-)-independent K+ pathway were less dependent on saline composition, and were not inhibited by amiloride, ethylisopropylamiloride, dimethylamiloride or gadolinium. Na+-K+-2Cl- cotransporter was inhibited whilst there was little effect on the Gardos channel (Ca2+-activated K+ channel). Peroxynitrite was markedly more effective in oxygenated cells than deoxygenated ones. Treatment with peroxynitrite per se did not affect initial cell volume. Anisotonic swelling modestly increased the Cl(-)-independent K+ influx, but did not affect peroxynitrite-stimulated KCC. Decreasing extracellular pH from 7.4 to 7.2 or 7.0 increased KCC stimulation, whilst the Cl(-)-independent component of K+ transport was lowest at pH 7.2. Finally, protein phosphatase inhibition with calyculin A (100 nM) inhibited KCC, implying that, as with other KCC stimuli, peroxynitrite acts via decreased protein phosphorylation; pre-treatment with calyculin A also inhibited the Cl(-)-independent component of K+ transport. These findings are relevant to the actions of peroxynitrite in vivo. 相似文献
12.
Paul Golby Nicholas P. J. Cotton Mark Carver J. Baz Jackson 《Archives of microbiology》1992,157(2):125-130
Comparison of the rate of accumulation of 86Rb+ by intact cells of Rhodobacter capsulatus during short periods of illumination with the Rb+-dependent membrane ionic current measured by electrochromism supports the view that both activities reflect the operation of a low-affinity K+ transport system. In experiments performed under similar conditions the ratio of 86Rb+ uptake to charge uptake was approx. 1.0, suggesting that the transport system operates as a uniporter. The addition of NH
inf4
sup+
to a cell suspension led to an increase in membrane ionic current but failed to inhibit the accumulation of 86Rb+ during illumination. The presence of K+ and NH
inf4
sup+
inhibited the increase in cellular ATP levels at the onset of illumination. This effect was prevented by Cs+. The results are considered within the context of the hypothesis (Golby et al. Eur J Biochem 187: 589–597) that NH
inf4
sup+
can be transported by the K+ carrier and in the context of an alternative hypothesis that NH
inf4
sup+
increases the affinity of the K+ transport system for its natural substrate and for Rb+.Abbreviations pH
pH difference across the cytoplasmic membrane
-
electrical potential difference across the cytoplasmic membrane 相似文献
13.
Effects of long-term, subtotal inhibition of Na+-K+ transport, either by growth of cells in sublethal concentrations of ouabain or in low-K+ medium, are described for HeLa cells. After prolonged growth in 2 × 10?8 M ouabain, the total number of ouabain molecules bound per cell increases by as much as a factor of three, mostly due to internalization of the drug. There is only about a 20% increase in ouabain-binding sites on the plasma membrane, representing amodest induction of Na+, K+-ATPase. In contrast, after long-term growth in low K+ there can be a twofold or greater increase in ouabain binding per cell, and in this case the additional sites are located in the plasma membrane. The increase is reversible. To assess the corresponding transport changes, we have separately estimated the contributions of increased intracellular [Na+] and of transport capacity (number of transport sites) to transport regulation. During both induction and reversal, short-term regulation is achieved primarily by changes in [Na+]i. More slowly, long-term regulation is achieved by changes in the number of functional transporters in the plasma membrane as assessed by ouabain binding, Vmax for transport, and specific phosphorylation. Parallel exposure of cryptic Na+, K+-ATPase activity with sodium dodecyl sulfate in the plasma membranes of both induced and control cells showed that the induction cannot be accounted for by an exposure of preexisting Na+, K+-ATPase in the plasma membrane. Analysis of the kinetics of reversal indicates that it may be due to a post-translational event. 相似文献
14.
Summary Hyperpolarization of voltage-clampedParamecium tetraurelia in K+ solutions elicits a complex of Ca2+ and K+ currents. The tail current that accompanies a return to holding potential (–40 mV) contains two K+ components. The tail current elicited by a step to –110 mV of 50-msec duration contains fast-decaying (3.5 msec) and slow-decaying (20 msec) components. The reversal potential of both components shifts by 55–57 mV/10-fold change in external [K+], suggesting that they represent pure K+ currents. The dependence of the relative amplitudes of the two tail currents on duration of hyperpolarization suggests that the slow K+ current activates slowly and is sustained, whereas the fast current activates rapidly during hyperpolarization and then rapidly inactivates. Iontophoretic injection of a Ca2+ chelator, EGTA, specifically reduces slow tail-current amplitude without affecting the fast tail component. Both K+ currents are inhibited by extracellular TEA+ in a concentration-dependent, noncooperative manner, whereas the fast K+ current alone is inhibited by 0.7mm quinidine. 相似文献
15.
A study was made of the ouabain effect (10(-3] on cell proliferation and the dependence of ATP hydrolysis on Na/K-concentration in homogenates of mouse hepatoma (XXIIa) and of L-cells, both sensitive and resistant to etidium bromide. Na+, K+-ATPase activity was found in homogenates of cells from sparse cultures in the presence of ouabain, the activity being stimulated by the Na/K-ratio pecular for the maximum enzymatic activity in cells from the dense cultures. The effect of ouabain on the cell proliferation is similar to the effect of transition of sparse cultures to dense ones. 相似文献
16.
17.
18.
19.
20.
Horst Helbig Christoph Korbmacher Michael Wiederholt 《The Journal of membrane biology》1987,99(3):173-186
Summary Using intracellular microelectrode technique, we investigated the changes in membrane voltage (V) of cultured bovine pigmented ciliary epithelial cells induced by different extracellular solutions. (1)V in 213 cells under steady-state conditions averaged –46.1±0.6 mV (sem). (2) Increasing extracellular K+ concentration ([K+]
o
) depolarizedV. Addition of Ba2+ could diminish this response. (3) Depolarization on doubling [K+]
o
was increased at higher [K+]
o
(or low voltage). (4) Removing extracellular Ca2+ decreasedV and reduced theV amplitude on increasing [K+]
o
. (5)V was pH sensitive. Extra-and intracellular acidification depolarizedV; alkalinization induced a hyperpolarization.V responses to high [K+]
o
were reduced at acidic extracellular pH. (6) Removing K
o
+
depolarized, K
o
+
readdition after K+ depletion transiently hyperpolarizedV. These responses were insensitive to Ba2+ but were abolished in the presence of ouabain or in Na+-free medium. (7) Na+ readdition after Na+ depletion transiently hyperpolarizedV. This reaction was markedly reduced in the presence of ouabain or in K+-free solution but unchanged by Ba2+. It is concluded that in cultured bovine pigmented ciliary epithelial cells K+ conductance depends on Ca2+, pH and [K+]
o
(or voltage). An electrogenic Na+/K+-transport is present, which is stimulated during recovery from K+ or Na+ depletion. This transport is inhibited by ouabain and in K+-or Na+-free medium. 相似文献