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1.
K+-conductive pathways were evaluated in isolated surface and crypt colonic cells, by measuring 86Rb efflux. In crypt cells, basal K+ efflux (rate constant: 0.24 ± 0.044 min−1, span: 24 ± 1.3%) was inhibited by 30 mM TEA and 5 mM Ba2+ in an additive way, suggesting the existence of two different conductive pathways. Basal efflux was insensitive to apamin, iberiotoxin, charybdotoxin and clotrimazole. Ionomycin (5 μM) stimulated K+ efflux, increasing the rate constant to 0.65 ± 0.007 min−1 and the span to 83 ± 3.2%. Ionomycin-induced K+ efflux was inhibited by clotrimazole (IC50 of 25 ± 0.4 μM) and charybdotoxin (IC50 of 65 ± 5.0 nM) and was insensitive to TEA, Ba2+, apamin and iberiotoxin, suggesting that this conductive pathway is related to the Ca2+-activated intermediate-conductance K+ channels (IKca). Absence of extracellular Ca2+ did neither affect basal nor ionomycin-induced K+ efflux. However, intracellular Ca2+ depletion totally inhibited the ionomycin-induced K+ efflux, indicating that the activation of these K+ channels mainly depends on intracellular calcium liberation. K+ efflux was stimulated by intracellular Ca2+ with an EC50 of 1.1 ± 0.04 μM. In surface cells, K+ efflux (rate constant: 0.17 ± 0.027 min−1; span: 25 ± 3.4%) was insensitive to TEA and Ba2+. However, ionomycin induced K+ efflux with characteristics identical to that observed in crypt cells. In conclusion, both surface and crypt cells present IKCa channels but only crypt cells have TEA- and Ba2+-sensitive conductive pathways, which would determine their participation in colonic K+ secretion.  相似文献   

2.
Summary To study the physiological role of the bidirectionally operating, furosemide-sensitive Na+/K+ transport system of human erythrocytes, the effect of furosemide on red cell cation and hemoglobin content was determined in cells incubated for 24 hr with ouabain in 145mm NaCl media containing 0 to 10mm K+ or Rb+. In pure Na+ media, furosemide accelerated cell Na+ gain and retarded cellular K+ loss. External K+ (5mm) had an effect similar to furosemide and markedly reduced the action of the drug on cellular cation content. External Rb+ accelerated the Na+ gain like K+, but did not affect the K+ retention induced by furosemide. The data are interpreted to indicate that the furosemide-sensitive Na+/K+ transport system of human erythrocytes mediates an equimolar extrusion of Na+ and K+ in Na+ media (Na+/K+ cotransport), a 1:1 K+/K+ (K+/Rb+) and Na+/Na+ exchange progressively appearing upon increasing external K+ (Rb+) concentrations to 5mm. The effect of furosemide (or external K+/Rb+) on cation contents was associated with a prevention of the cell shrinkage seen in pure Na+ media, or with a cell swelling, indicating that the furosemide-sensitive Na+/K+ transport system is involved in the control of cell volume of human erythrocytes. The action of furosemide on cellular volume and cation content tended to disappear at 5mm external K+ or Rb+. Thein vivo red cell K+ content was negatively correlated to the rate of furosemide-sensitive K+ (Rb+) uptake, and a positive correlation was seen between mean cellular hemoglobin content and furosemide-sensitive transport activity. The transport system possibly functions as a K+ and waterextruding mechanism under physiological conditiosin vivo. The red cell Na+ content showed no correlation to the activity of the furosemide-sensitive transport system.  相似文献   

3.
Summary The effects of cAMP, ATP and GTP on the Ca2+-dependent K+ channel of fresh (1–2 days) or cold-stored (28–36 days) human red cells were studied using atomic absorption flame photometry of Ca2+-EGTA loaded ghosts which had been resealed to monovalent cations in dextran solutions. When high-K+ ghosts were incubated in an isotonic Na+ medium, the rate constant of Ca2+-dependent K+ efflux was reduced by a half on increasing the theophylline concentration to 40mm. This effect was observed in ghosts from both fresh and stored cells, but only if they were previously loaded with ATP. The inhibition was more marked when Mg2+ was added together with ATP, and it was abolished by raising free Ca2+ to the micromolar level. Like theophylline, isobutyl methylxanthine (10mm) also affected K+ efflux. cAMP (0.2–0.5mm), added both internally and externally (as free salt, dibutyryl or bromide derivatives), had no significant effect on K+ loss when the ghost free-Ca2+ level was below 1 m, but it was slightly inhibitory at higher concentrations. The combined presence of cAMP (0.2mm) plus either theophylline (10mm), or isobutyl methylxanthine (0.5mm), was more effective than cAMP alone. This inhibition showed a strict requirement for ATP plus Mg2+ and it, was not overcome by raising internal Ca2+. Ghosts from stored cells seemed more sensitive than those from fresh cells, to the combined action of cAMP and methylxanthines. Loading ATP into ghosts from fresh or stored cells markedly decreased K+ loss. Although this effect was observed in the absence of added Mg2+ (0.5mm EDTA present), it was potentiated upon adding 2mm Mg2+. The K+ efflux from ATP-loaded ghosts was not altered by dithio-bis-nitrobenzoic acid (10mm) or acridine orange (100 m), while it was increased two-to fourfold by incubating with MgF2 (10mm), or MgF2 (10mm)+theophylline (40mm), respectively. By contrast, a marked efflux reduction was obtained by incorporating 0.5mm GTP into ATP-containing ghosts. The degree of phosphorylation obtained by incubating membranes with (-32P)ATP under various conditions affecting K+ channel activity, was in direct correspondence to their effect on K+ efflux. The results suggest that the K+ channel of red cells is under complex metabolic control, via cAMP-mediated and nonmediated mechanisms, some which require ATP and presumably, involve phosphorylation of the channel proteins.  相似文献   

4.
Summary The apical membrane K+ permeability of the newt proximal tubular cells was examined in the doubly perfused isolated kidney by measuring the apical membrane potential change (V a change) during alteration of luminal K+ concentration and resultant voltage deflections caused by current pulse injection into the lumen.V a change/decade for K+ was 50 mV at K+ concentration higher than 25mm, and the resistance of the apical membrane decreased bt 58% of control when luminal K+ concentration was increased from 2.5 to 25mm. Ba2+ (1mm in the lumen) reducedV a change/decade to 24 mV and increased the apical membrane resistance by 70%. These data support the view that Ba2+-sensitive K+ conductance exists in the apical membrane of the newt proximal tubule. Furthermore, intracellular K+ activity measured by K+-selective electrode was 82.4 ± 3.6 meq/liter, which was higher than that predicted from the Nernst equation for K+ across both cell membranes. Thus, it is concluded that cell K+ passively diffuses, at least in part, through the K+ conductive pathway of the apical membrane.  相似文献   

5.
Palytoxin (PTX) opens a pathway for ions to pass through Na,K-ATPase. We investigate here whether PTX also acts on nongastric H,K-ATPases. The following combinations of cRNA were expressed in Xenopus laevis oocytes: Bufo marinus bladder H,K-ATPase α2- and Na,K-ATPase β2-subunits; Bufo Na,K-ATPase α1- and Na,K-ATPase β2-subunits; and Bufo Na,K-ATPase β2-subunit alone. The response to PTX was measured after blocking endogenous Xenopus Na,K-ATPase with 10 μm ouabain. Functional expression was confirmed by measuring 86Rb uptake. PTX (5 nm) produced a large increase of membrane conductance in oocytes expressing Bufo Na,K-ATPase, but no significant increase occurred in oocytes expressing Bufo H,K-ATPase or in those injected with Bufo β2-subunit alone. Expression of the following combinations of cDNA was investigated in HeLa cells: rat colonic H,K-ATPase α1-subunit and Na,K-ATPase β1-subunit; rat Na,K-ATPase α2-subunit and Na,K-ATPase β2-subunit; and rat Na,K-ATPase β1- or Na,K-ATPase β2-subunit alone. Measurement of increases in 86Rb uptake confirmed that both rat Na,K and H,K pumps were functional in HeLa cells expressing rat colonic HKα1/NKβ1 and NKα2/NKβ2. Whole-cell patch-clamp measurements in HeLa cells expressing rat colonic HKα1/NKβ1 exposed to 100 nm PTX showed no significant increase of membrane current, and there was no membrane conductance increase in HeLa cells transfected with rat NKβ1- or rat NKβ2-subunit alone. However, in HeLa cells expressing rat NKα2/NKβ2, outward current was observed after pump activation by 20 mm K+ and a large membrane conductance increase occurred after 100 nm PTX. We conclude that nongastric H,K-ATPases are not sensitive to PTX when expressed in these cells, whereas PTX does act on Na,K-ATPase.  相似文献   

6.
Properties of potassium uptake by seedling roots of grape cultivars   总被引:1,自引:0,他引:1  
Uptake rates of (86Rb)K+ by seedling roots of six cultivars were measured and compared with K+ content of the root, K+ leakage, H+ efflux, and K+-ATPase activity of a partially purified plasmalemma fraction.Different cultivars showed significantly different rates of (86Rb)K+ uptake. The uptake rates of the first (0–5 min) period did not correlate with K+ content of the seedling roots.The rates of uptake in the 10 to 30 min period, supposed to be active, were negatively correlated with K+ content of the root. Roots consistently leaked K+ during the first 5 min. This leakage was positively correlated with the endogenous K+ content of the tissue.H+ efflux was significantly different among the cultivars and correlated with the K+-ATPase activity of a microsomal fraction partially purified on discontinuous (18/34%) sucrose gradient. The relationships among transport parameters are discussed.Part of the present research was carried out at the Department of Agricultural Biotechnology of University of Padova, Italy.Part of the present research was carried out at the Department of Agricultural Biotechnology of University of Padova, Italy.  相似文献   

7.
Hickey KD  Buhr MM 《Theriogenology》2012,77(7):1369-1380
Existing as a ubiquitous transmembrane protein, Na+K+-ATPase affects sperm fertility and capacitation through ion transport and a recently identified signaling function. Functional Na+K+-ATPase is a dimer of α and β subunits, each with isoforms (four and three, respectively). Since specific isoform pairings and locations may influence or indicate function, the objective of this study was to identify and localize subunits of Na+K+-ATPase in fresh bull sperm by immunoblotting and immunocytochemistry using antibodies against α1 and 3, and all β isoforms. Relative quantity of Na+K+-ATPase in head plasma membranes (HPM's) from sperm of different bulls was determined by densitometry of immunoblot bands, and compared to bovine kidney. Sperm and kidney specifically bound all antibodies at kDa equivalent to commercial controls, and to additional lower kDa bands in HPM. Immunofluorescence of intact sperm confirmed that all isoforms were present in the head region of sperm and that α3 was also uniformly distributed post-equatorially. Permeabilization exposing internal membranes typically resulted in an increase in fluorescence, indicating that some antibody binding sites were present on the inner surface of the HPM or the acrosomal membrane. Deglycosylation of β1 reduced the kDa of bands in sperm, rat brain and kidney, with the kDa of the deglycosylated bands differing among tissues. Two-dimensional blots of β1 revealed three distinct spots. Based on the unique quantity, location and structure Na+K+-ATPase subunits in sperm, we inferred that this protein has unique functions in sperm.  相似文献   

8.
Summary Transbasal electrical potential (V b) and intraepithelial potassium chemical activity ((K+) i ) were measured in isolated midgut epithelium of tobacco hornworm (Manduca sexta) using double-barrelled glass microelectrodes. Values ofV b ranging from +8 to –48 mV (relative to blood side) were recorded. For all sites, (K+) i is within a few millivolts of electrochemical equilibrium with the blood side bathing solution. Sites more negative than –20 mV show relatively high sensitivity ofV b to changes in blood side K+ concentration: 43% of these sites can be marked successfully with iontophoresed Lucifer yellow CH dye and shown to represent epithelial cells of all three types present in the midgut. In about half of successful marks, dye-coupling of several adjacent cells is seen. Low potential sites — those withV b less negative than –20 mV —typically do not show high sensitivity ofVb to changes of external K+, but rather (K+) i rapidly approaches the K+ activity of blood side bathing solution. These sites can seldom be marked with Lucifer yellow (4% success). The mean (K+) i of the high potential sites is 95±29 (sd)mm under standard conditions, a value which is in accord with published values for the whole tissue.  相似文献   

9.
Summary Chronic exposure to high potassium stimulates K+-secretory mechanisms in the diluting segment of the amphibian kidney (K+ adaptation). Since K+ net flux depends critically on the passive cell membrane permeabilities for K+ ions, cable analysis and K+-concentration step changes were applied in this nephron segment to assess the individual resistances of the epithelium and the K+ conductance of the luminal cell membrane. Experiments were performed in the isolated, doubly-perfused kidney of both control and K+-adaptedAmphiuma. In control animals transepithelial resistance was 290±27 cm2, which decreased significantly to 199±17 cm2 after K+ adaptation. The resistance in parallel of the luminal and peritubular cell membrane decreased from a control value of 157±14 to 108±6 cm2 after chronic K+ treatment. This was paralleled by a decrease of the ratio of the luminal to peritubular cell membrane resistance from 2.5±0.1 to 1.9±0.1, respectively. Estimation of the individual cell membrane resistances reveals that the combined resistance of the luminal and peritubular cell membrane is in the same order of magnitude as the paracellular shunt resistance in diluting segments of both control and K+-adapted animals. The luminal cell membrane is K+ selective under both conditions, but the absolute luminal K+ conductance increases by some 60% with K+ adaptation. This leads to an increased back-leak of K+ from cell to lumen and may explain stimulated K+ net secretion found after chronic K+ loading.  相似文献   

10.
Regulation of the Na+/K+-ATPase by insulin: Why and how?   总被引:4,自引:0,他引:4  
The sodium-potassium ATPase (Na+/K+-ATPase or Na+/K+-pump) is an enzyme present at the surface of all eukaryotic cells, which actively extrudes Na+ from cells in exchange for K+ at a ratio of 3:2, respectively. Its activity also provides the driving force for secondary active transport of solutes such as amino acids, phosphate, vitamins and, in epithelial cells, glucose. The enzyme consists of two subunits ( and ) each expressed in several isoforms. Many hormones regulate Na+/K+ -ATPase activity and in this review we will focus on the effects of insulin. The possible mechanisms whereby insulin controls Na+/K+-ATPase activity are discussed. These are tissue- and isoform-specific, and include reversible covalent modification of catalytic subunits, activation by a rise in intracellular Na+ concentration, altered Na+ sensitivity and changes in subunit gene or protein expression. Given the recent escalation in knowledge of insulin-stimulated signal transduction systems, it is pertinent to ask which intracellular signalling pathways are utilized by insulin in controlling Na+/K+-ATPase activity. Evidence for and against a role for the phosphatidylinositol-3-kinase and mitogen activated protein kinase arms of the insulin-stimulated intracellular signalling networks is suggested. Finally, the clinical relevance of Na+/K+-ATPase control by insulin in diabetes and related disorders is addressed.  相似文献   

11.
Na+, K+-pumps of most eukaryotic animal cells bind ouabain with high affinity, stop pumping, and consequently loose K+, detach from each other and from the substrate, and die. Lack of affinity for the drug results in ouabain resistance. In this work, we report that Ma104 cells (epithelial from Rhesus monkey kidney) have a novel form of ouabain-resistance: they bind the drug with high affinity (Km about 4×10–8 m), they loose their K+ and stop proliferating but, in spite of these, up to 100% of the cells remain attached in 1.0 m ouabain, and 53% in 1.0 mm. When 4 days later ouabain is removed from the culture medium, cells regain K+ and resume proliferation. Strophanthidin, a drug that attaches less firmly than ouabain, produces a similar phenomenon, but allows a considerably faster recovery. This reversal may be associated to the fact that, while in ouabain-sensitive MDCK cells Na+, K+-ATPases blocked by the drug are retrieved from the plasma membrane, those in Ma104 cells remain at the cell-cell border, as if they were cell-cell attaching molecules. Cycloheximide (10 g/ml) and chloroquine (10 m) impair this recovery, suggesting that it also depends on the synthesis and insertion of a crucial protein component, that may be different from the pump itself. Therefore ouabain resistance of Ma104 cells is not due to a lack of affinity for the drug, but to a failure of its Na+, K+-ATPases to detach from the plasma membrane in spite of being blocked by ouabain.We wish to thank Dr. E. Rodríguez-Boulán for the generous supply of Ma104 cells, as well as acknowledge the generous economic support of the National Research Council (CONACYT) of Mexico. Confocal experiments were performed in the Confocal Microscopy Unit of the Physiology Department, CINVESTAV.  相似文献   

12.
The effect of point mutation in the sequence 316TWLE319, which occurs in the extracellular loop flanking the third (M3) and the fourth (M4) transmembrane segment (L3/4) of the Na+,K+-ATPase alpha-subunit, was examined. Mutation of Glu319 to Asp yielded an enzyme with full activity, whereas substituting Glu319 to Ala resulted in a severe loss of activity. A negative charge was introduced along the sequence, one residue at a time, from Thr316 to Leu318 (by E-scanning) in the mutant construct with Glu319 already mutated to Gln. The activity that had been reduced to 60% by the mutation of Glu319 to Gln was restored upon the introduction of a negative charge by E-scanning. When Leu318 was replaced by Glu in a series of scanning experiments, the K+ sensitivity of the ATPase activity was lowered. The lowering of K+ sensitivity was further demonstrated when a mutation of Leu318 to Glu was introduced into the wild-type enzyme. Furthermore, mutants with Leu318 to Gln, Arg, and Phe displayed lower K+ sensitivity similar to that of Leu318 to Glu mutant. Leu318 may be in access path for K+, and any substitution at this position may interfere with access of K+ from outside the cell.  相似文献   

13.
The Shaker B K(+) conductance (G(K)) collapses (in a reversible manner) if the membrane is depolarized and then repolarized in, 0 K(+), Na(+)-containing solutions (Gómez-Lagunas, F. 1997. J. Physiol. 499:3-15; Gómez-Lagunas, F. 1999. Biophys. J. 77:2988-2998). In this work, the role of Na(+) ions in the collapse of G(K) in 0-K(+) solutions, and in the behavior of the channels in low K(+) was studied. The main findings are as follows. First, in 0-K(+) solutions, the presence of Na(+) ions is an important factor that speeds the collapse of G(K). Second, external Na(+) fosters the drop of G(K) by binding to a site with a K(d) = 3.3 mM. External K(+) competes, in a mutually exclusive manner, with Na(o)(+) for binding to this site, with an estimated K(d) = 80 microM. Third, NMG and choline are relatively inert regarding the stability of G(K); fourth, with [K(o)(+)] = 0, the energy required to relieve Na(i)(+) block of Shaker (French, R.J., and J.B. Wells. 1977. J. Gen. Physiol. 70:707-724; Starkus, J.G., L. Kuschel, M. Rayner, and S. Heinemann. 2000. J. Gen. Physiol. 110:539-550) decreases with the molar fraction of Na(i)(+) (X(Na,i)), in an extent not accounted for by the change in Delta(mu)(Na). Finally, when X(Na,i) = 1, G(K) collapses by the binding of Na(i)(+) to two sites, with apparent K(d)s of 2 and 14.3 mM.  相似文献   

14.
The insoluble fraction of ox-brain, which had previously been shown to have a non-linear affinity for Na+ and K+, was prepared. Acetylcholine (1×10–8 mol/l and 1×10–7 mol/l) reduced the affinity of the fraction for Na+ and K+ to zero, while at 1×10–6 mol/l, the affinity for the cations was almost as high as in the absence of the transmitter; the affinities for Na+ and K+ were particularly high, when the supernatant concentrations of these ions exceeded 80–100 mM. Addition of eserine (3×10–5 mol/l) considerably modified the response of the fraction to acetylcholine (1×10–5 mol/l). Atropine (1×10–8 mol/l) in the absence or presence of acetylcholine (1×10–5, or 1×10–4 mol/l) reduced the affinity of the fraction for Na+ and K+ to zero. Epinephrine (3×10–10 mol/l) lowered the affinity for Na+ and K+, while ergotamine itself (1×10–5 mol/l) reduced it to zero. The addition of both epinephrine and ergotamine at the latter concentrations restored the affinity of the fractions for Na+ and K+ to what it had been in the absence of the transmitter or antagonist, previously reported. Norepinephrine (3×10–10 mol/l), or ouabain (1×10–7 mol/l) reduced the affinity of the fraction for Na+ and K+ to zero. Thus, the transmitters and antagonists altered the affinity of the insoluble fraction for Na+ and K+ nonlinearity, dependent upon their concentrations, the concentrations of the cations, and the interaction of transmitter and antagonist.  相似文献   

15.
Leucines were mutated within the sequence L311ILGYTWLE319 of the extracellular loop flanking the third (M3) and fourth (M4) transmembrane segments (M3/M4 loop) of the Torpedo Na+,K+-ATPase α-subunit. Replacement of Leu311 with Glu resulted in a considerable loss of Na+,K+-ATPase activity. Replacement of Leu313 with Glu shifted the equilibrium of E1P and E2P toward E1P and reduced the rate of the E1P to E2P transition. The reduction of the transition rate and stronger inhibition of Na+,K+-ATPase activity by Na+ at higher concentrations together suggest that there is interference of Na+ release on the extracellular side in the Leu313 mutant. Thus, Leu313 could be in the pathway of Na+ exit. Replacement of Leu318 with Glu yielded an enzyme with significantly reduced apparent affinity for both vanadate and K+, with an equilibrium shifted toward E2P and no alteration in the transition rate. The reduced vanadate affinity is due to the lower rate of production of vanadate-reactive [K+ 2]E2 caused by inhibition of dephosphorylation through reduction of the K+ affinity of E2P. Thus, Leu318 may be a critical position in guiding external K+ to its binding site.  相似文献   

16.
Alves DP  Tatsuo MA  Leite R  Duarte ID 《Life sciences》2004,74(20):2577-2591
In order to investigate to the contribution of K+ channels on the peripheral antinociception induced by diclofenac, we evaluated the effect of several K+ channel blockers, using the rat paw pressure test, in which sensitivity is increased by intraplantar injection (2 microg) of prostaglandin E2. Diclofenac administered locally into the right hindpaw (25, 50, 100 and 200 microg) elicited a dose-dependent antinociceptive effect which was demonstrated to be local, since only higher doses produced an effect when injected in the contralateral paw. This blockade of PGE2 mechanical hyperalgesia induced by diclofenac (100 microg/paw) was antagonized in a dose-dependent manner by intraplantar administration of the sulphonylureas glibenclamide (40, 80 and 160 microg) and tolbutamide (80, 160 and 320 microg), specific blockers of ATP-sensitive K+ channels, and it was observed even when the hyperalgesic agent used was carrageenin, while the antinociceptive action of indomethacin (200 microg/paw), a typical cyclo-oxygenase inhibitor, over carrageenin-induced hyperalgesia was not affected by this treatment. Charybdotoxin (2 microg/paw), a blocker of large conductance Ca2+-activated K+ channels and dequalinium (50 microg/paw), a selective blocker of small conductance Ca2+-activated K+ channels, did not modify the effect of diclofenac. This effect was also unaffected by intraplantar administration of non-specific voltage-dependent K+ channel blockers tetraethylammonium (1700 microg) and 4-aminopyridine (100 microg) or cesium (500 microg), a non-specific K+ channel blocker. The peripheral antinociceptive effect induced by diclofenac was antagonized by NG-Nitro L-arginine (NOarg, 50 microg/paw), a NO synthase inhibitor and methylene blue (MB, 500 microg/paw), a guanylate cyclase inhibitor, and this antagonism was reversed by diazoxide (300 microg/paw), an ATP-sensitive K+ channel opener. We also suggest that an endogenous opioid system may not be involved since naloxone (50 microg/paw) did not affect diclofenac-induced antinociception in the PGE2-induced hyperalgesia model. This study provides evidence that the peripheral antinociceptive effect of diclofenac may result from activation of ATP-sensitive K+ channels, possible involving stimulation of L-arginine/NO/cGMP pathway, while Ca2+-activated K+ channels, voltage-dependent K+ channels as well as endogenous opioids appear not to be involved in the process.  相似文献   

17.
H+-ATPase activity of a plasma membrane-enriched fraction decreased after the treatment of barley (Hordeum vulgare) seedlings with Al for 5 days. A remarkably high level of Al was found in the membrane fraction of Al-treated roots. A long-term effect of Al was identified as the repression of the H+-ATPase of plasma membranes isolated from the roots of barley and wheat (Triticum aestivum) cultivars, Atlas 66 (Al-tolerant) and Scout 66 (Al-sensitive). To monitor short-term effects of Al, the electrical membrane potentials across plasma membranes of both wheat cultivars were compared indirectly by measuring the efflux of K+ for 40 min under various conditions. The rate of efflux of K+ in Scout was twice that in Atlas at low pH values such as 4.2. Vanadate, an inhibitor of the H+-ATPase of the plasma membrane, increased the efflux of K+. Al repressed this efflux at low pH, probably through an effect on K+ channels, and repression was more pronounced in Scout. Al strongly repressed the efflux of K+ irrespective of the presence of vanadate. Ca2+ also had a repressive effect on the efflux of K+ at low pH. The effect of Ca2+, greater in Scout, might be related to the regulation of the net influx of H+, since the effect was negated by vanadate. The results suggest that extracellular low pH may cause an increase in the influx of H+, which in turn is counteracted by the efflux of K+ and H+. These results suggest that the ability to maintain the integrity of the plasma membrane and the ability to recover the electrical balance at the plasma membrane through a net influx of H+ and the efflux of K+ seem to participate in the mechanism of tolerance to Al stress under acidic conditions.  相似文献   

18.
It is reported that CNS hemorrage causes membrane dysfunction and may exacerbate this damage as a result of secondary ischemia or hypoxia. Since hyperbaric oxygenation improves oxygen metabolism, it may reduce this membrane damage. The present study was conducted to reveal whether hyperbaric oxygenation influences membrane alteration after hemorrhage. Thirty minutes after subarachnoid hemorrhage induction, rats were treated with hyperbaric oxygenation 2 ATA for 1 hour. Rats were decapitated 2 hours after subarachnoid hemorrhage induction. Na+, K+-ATPase activity measurement, and spin-label studies were performed on crude synpatosomal membranes. Subarachnoid hemorrhage decreased Na+, K+-ATPase activity. Spin label studies showed that hydrophobic portions of near the membrane surface became more rigid and the mobility of the membrane protein labeled sulfhydryl groups decreased after subarachnoid hemorrhage. Hyperbaric oxygenation significantly ameliorated most of the subarachnoid hemorrhage induced alterations. We conclude that hyperbaric oxygenation may be a beneficial treatment for acute subarachnoid hemorrhage.  相似文献   

19.
Nonesterified long-chain fatty acids (myristic, palmitic, oleic and arachidonic), added at low amounts (around 20 nmol/mg protein) to rat liver mitochondria, energized by respiratory substrates and suspended in isotonic solutions of KCl, NaCl, RbCl or CsCl, adjusted to pH 8.0, induce a large-scale swelling followed by a spontaneous contraction. Such swelling does not occur in alkaline solutions of choline chloride or potassium gluconate or sucrose. These changes in the matrix volume reflect a net uptake, followed by net extrusion, of KCl (or another alkali metal chloride) and are characterized by the following features: (1) Lowering of medium pH from 8.0 to 7.2 results in a disappearance of the swelling-contraction reaction. (2) The contraction phase disappears when the respiration is blocked by antimycin A. (3) Quinine, an inhibitor of the K+/H+ antiporter, does not affect swelling but suppresses the contraction phase. (4) The swelling phase is accompanied by a decrease of the transmembrane potential and an increase of respiration, whereas the contraction is followed by an increase of the membrane potential and a decrease of oxygen uptake. (5) Nigericin, a catalyst of the K+/H+ exchange, prevents or partly reverses the swelling and partly restores the depressed membrane potential. These results indicate that long-chain fatty acids activate in liver mitochondria suspended in alkaline saline media the uniporter of monovalent alkali metal cations, the K+/H+ antiporter and the inner membrane anion channel. These effects are presumably related to depletion of mitochondrial Mg2+, as reported previously [Arch. Biochem. Biophys. 403 (2002) 16], and are responsible for the energy-dissipating K+ cycling. The uniporter and the K+/H+ antiporter are in different ways activated by membrane stretching and/or unfolding, resulting in swelling followed by contraction.  相似文献   

20.
Summary The initial rate of ATP-dependent proton uptake by hog gastric vesicles was measured at pH's between 6.1 and 6.9 by measuring the loss of protons from the external space with a glass electrode. The apparent rates of proton loss were corrected for scalar proton production due to ATP hydrolysis. For vesicles in 150mm KCl and pH 6.1, corrected rates of proton uptake and ATP hydrolysis were 639±84 and 619±65 nmol/min×mg protein, respectively, giving an H+/ATP ratio of 1.03±0.7. Furthermore, at all pH's tested the ratio of the rate of proton uptake to the rate of ATP hydrolysis was not significantly different than 1.0. No proton uptake (<10 nmol/min×mg protein) was exhibited by vesicles in 150mm NaCl at pH 6.1 despite ATP hydrolysis of 187±46 nmol/min×mg (nonproductive hydrolysis). Comparison of the rates of proton transport and ATP hydrolysis in various mixture of KCl and NaCl showed that the H+/ATP stoichiometries were not significantly different than 1.0 at all concentrations of K+ greater than 10mm. This fact suggests that the nonproductive rate is vanishingly small at these concentrations, implying that the measured H+/ATP stoichiometry is equal to the enzymatic stoichiometry. This result shows that the isolated gastric (K++H+)-ATPase is thermodynamically capable of forming the observed proton gradient of the stomach.  相似文献   

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