首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 31 毫秒
1.
The uptake of K+ and Ca2+ in Dunaliella salina is mediated by two distinct carriers: a K+ carrier with a high selectivity against Na+, Li+, and choline+ but not towards Rb+, K+, Cs+, or NH4+, and a Ca2+ carrier with a high selectivity against Mg2+. The latter is specifically blocked by La3+ and by Cd2+. Apparent Km values for K+ and Ca2+ uptake are 2.5 and 0.8 millimolar, respectively, and their maximal calculated fluxes are 22 and 0.8 nanomoles per square meter per second, respectively. Effects of permeable ions and ionophores on K+ and Ca2+ uptake suggest that the driving force for their uptake is the transmembrane electrical potential. Inhibitors of ATP production, typical inhibitors of plasma membrane H+-ATPases and protonionophores inhibit K+ and Ca2+ uptake and accelerate K+ efflux. The results suggest that an H+-ATPase in the cell membrane provides the driving force for K+ and Ca2+ uptake. Efflux measurements from 86Rb+ and 45Ca2+ loaded cells suggest that part of the intracellular K+ and most of the intracellular Ca2+ is nonexchangeable with the extracellular pool. Correlations between phosphate and K+ contents and the effect of phosphate on K+ efflux suggest intracellular associations between K+ and polyphosphates. On the basis of these results, it is suggested that: (a) K+ and Ca2+ uptake in D. salina is driven by the transmembrane electrical potential which is generated by the action of an H+-ATPase of the plasma membrane. (b) Part of the intracellular K+ is associated with polyphosphate bodies, while most of the intracellular Ca2+ is accumulated in intracellular organelles in the algal cells.  相似文献   

2.
Three cultivars of sugar beet (Beta vulgaris L.), which are sensitive to aluminium (Al) in the order Primahill > Monohill > Regina, were grown in water culture for 2 weeks. Nutrients were supplied at 15% increase of amounts daily, corresponding to the nutrient demand for maximal growth. The 2.4-dinitrophenol (DNP)-sensitive (metabolic) and DNP-insensitive (non-metabolic) uptake of aluminium, phosphate. 45Ca2+ and K+(86Rb+) in roots were measured as well as transport to shoots of intact plants. All 3 cultivars absorbed more aluminium if DNP was present during the aluminium treatment than in its absence. It is suggested that sugar beets are able to extrude aluminium activity or that they possess an active mechanism to keep Al outside the cell. The presence of Al in the medium during the 1-h experiment affected the metabolic and non-metabolic fluxes of 45Ca2+ and K+(86Rb+) in different ways. In the presence of DNP, the influx of both 45Ca2+ and K+(86Rb+) and the efflux of 45Ca2+ were inhibited by Al in a competitive way. At inhibition of 45Ca2+ influx, 2 Al ions are probably bound per Ca2+ uptake site in cv. Regina (Al-tolerant), but in cvs Primahill and Monohill only one Al ion is bound (more Al sensitive). Aluminium competitively inhibited the active efflux of 45Ca2+ (absence of DNP) in almost the same way in the 3 cultivars. In contrast, aluminium stimulated the influx of K+(86Rb+) in cvs Primahill, Monohill and Regina in the absence of DNP. Thus, the Al effects on active and passive K+(86Rb+) influx are different. The total influx of K+(86Rb+) increased in the presence of Al and might be connected to an active exclusion of Al. Regina is the least Al-sensitive cultivar, probably because Al interferes less with the Ca2+ fluxes and because this cultivar actively excludes phosphate in the presence of Al. Thus Al-phosphate precipitation within the plant could be avoided.  相似文献   

3.
Oxidative stress to B-cells is thought to be of relevance in declining B-cell function and in the process of B-cell destruction. In other tissues including heart, brain and liver, oxidative stress has been shown to elevate the intracellular free calcium concentration and to provoke potassium efflux. We studied the effect of oxidative stress on Ca2+ and K+ (Rb+) outflow from pancreatic islets using the thiol oxidants DIP and BuOOH. Both compounds reversibly increased 86Rb+ efflux in the presence of 3 and 16.7 mmol/l glucose. Stimulation of 86Rb+ efflux was also evident in the absence of calcium. DIP evoked release of 45Ca2+ from the pancreatic islets both in the presence or absence of extracellular calcium. Employing inhibitors of the calcium-activated potassium channel (KCa) and the high conductance K+-channel (BKCa), the effect of DIP on 86Rb+ efflux was slightly diminished. Tolbutamide had no effect on 86Rb+ efflux in the presence of DIP. On the other hand thapsigargin, a blocker of the Ca2+-ATPase of the endoplasmic reticulum, completely suppressed the DIP-mediated 86Rb+ outflow. The data suggest that thiol oxidant-induced potassium efflux from pancreatic islets is mainly mediated through liberation of intracellular calcium and subsequent stimulation of calcium-activated potassium efflux.  相似文献   

4.
We have previously demonstrated mobilization of Ca2+ in the efflux of Rb+ (K+) from isolated hamster brown adipocytes as a consequence of norepinephrine stimulation. We have now investigated the adrenoceptor subtype specificity of these responses and found them both to be of theα1-subtype. Futher, we have found that the Rb+ (K+) effux was dependent upon a primary Ca2+mobilization event in response to the α1-adrenergic stimulation, since the Rb+ efflux could also be demonstrated by the addition ionophore A23187 to the cells. The norepinephrine- and A23187-stimulated Rb+ effluxes were both inhibited by the Ca2+-dependent K+ -channel blocker apamin. Apamin also significantly attenuated Ca2+ mobilization in cells in response to a submaximal concentration of norepinephrine. We conclude that α1-adrenergic stimulation of brown fat cells leads to a mobilization of intracellular Ca2+ which, in itself or via other mechanisms, leads to an increase in cytosolic Ca2+ concentration which, in turn, activates a Ca2+ -dependent K+ channel leading to a K+ release from these cells. A possible role for this channel to sustain and augment the response toα1-adrenergic stimulation is discussed.  相似文献   

5.
The effect of a hyposmotic shock and extracellular ATP on the efflux of K+(Rb+) from human breast cancer cell lines (MDA-MB-231 and MCF-7) has been examined. A hyposmotic shock increased the fractional efflux of K+(Rb+) from MDA-MB-231 cells via a pathway which was unaffected by Cl replacement. Apamin, charybdotoxin or removing extracellular Ca2+ had no effect on volume-activated K+(Rb+) efflux MDA-MB-231 cells. An osmotic shock also stimulated K+(Rb+) efflux from MCF-7 cells but to a much lesser extent than found with MDA-MB-231 cells. ATP-stimulated K+(Rb+) efflux from MDA-MB-231 cells in a dose-dependent fashion but had little effect on K+(Rb+) release from MCF-7 cells. ATP-stimulated K+(Rb+) efflux was only inhibited slightly by replacing Cl with NO3. Removal of external Ca2+ during treatment with ATP reduced the fractional efflux of K+(Rb+) in a manner suggesting a role for cellular Ca2+ stores. Charybdotoxin, but neither apamin nor iberiotoxin, inhibited ATP-stimulated K+(Rb+) release from MDA-MB-231 cells. Suramin inhibited the ATP-activated efflux of K+(Rb+). UTP also stimulated K+(Rb+) efflux from MDA-MB-231 cells whereas ADP, AMP and adenosine were without effect. A combination of an osmotic shock and ATP increased the fractional efflux of K+(Rb+) to a level greater than the sum of the individual treatments. It appears that the hyposmotically-activated and ATP-stimulated K+ efflux pathways are separate entities. However, there may be a degree of ‘crosstalk’ between the two pathways.  相似文献   

6.
Summary In the mammalian distal colon, the surface epithelium is responsible for electrolyte absorption, while the crypts are the site of secretion. This study examines the properties of electrical potential-driven86Rb+ fluxes through K+ channels in basolateral membrane vesicles of surface and crypt cells of the rabbit distal colon epithelium. We show that Ba2+-sensitive, Ca2+-activated K+ channels are present in both surface and crypt cell derived vesicles with half-maximal activation at 5×10–7 m free Ca2+. This suggests an important role of cytoplasmic Ca2+ in the regulation of the bidirectional ion fluxes in the colon epithelium.The properties of K+ channels in the surface cell membrane fraction differ from those of the channels in the crypt cell derived membranes. The peptide toxin apamin inhibits Ca2+-activated K+ channels exclusively in surface cell vesicles, while charybdotoxin inhibits predominantely in the crypt cell membrane fraction. Titrations with H+ and tetraethylammonium show that both high-and low-sensitive86Rb+ flux components are present in surface cell vesicles, while the high-sensitive component is absent in the crypt cell membrane fraction. The Ba2+-sensitive, Ca2+-activated K+ channels can be solubilized in CHAPS and reconstituted into phospholipid vesicles. This is an essential step for further characterization of channel properties and for identification of the channel proteins in purification procedures.  相似文献   

7.
Summary 86Rb+ fluxes have been measured in suspensions of vesicles prepared from the epithelium of toad urinary bladder. A readily measurable barium-sensitive, ouabain-insensitive component has been identified; the concentration of external Ba2+ required for half-maximal inhibition was 0.6mm. The effects of externally added cations on86Rb+ influx and efflux have established that this pathway is conductive, with a selectivity for K+, Rb+ and Cs+ over Na+ and Li+. the Rb+ uptake is inversely dependent on external pH, but not significantly affected by internal Ca2+ or external amiloride, quinine, quinidine or lidocaine. It is likely, albeit not yet certain, that the conductive Rb+ pathway is incorporated in basolateral vesicles oriented right-side-out. It is also not yet clear whether this pathway comprises the principle basolateral K+ channel in vivo, and that its properties have been unchanged during the preparative procedures. Subject to these caveats, the data suggest that the inhibition by quinidine of Na+ transport across toad bladder does not arise primarily from membrane depolarization produced by a direct blockage of the basolateral channels. It now seems more likely that the quinidine-induced elevation of intracellular Ca2+ activity directly blocks apical Na+ entry.  相似文献   

8.
The mechanism of the protective effect of Ca2+ on cellular K+ content was studied by examination of the effect of Ca2+ on efflux of the K+ analog, 86Rb+, from preloaded cells with the use of compounds which interfere with monovalent cation movements. Ca2+ decreased 86Rb+ efflux to the same extent in the presence and absence of ouabain, suggesting that Ca2+ did not alter the activity of the (Na+ + K+)-adenosine triphosphatase pump. Ca2+ exerted a similar protective effect in the presence of furosemide, an inhibitor of K+-K+ exchange, indicative that Ca2+ was not inhibiting this pathway. Since Ca2+ did not influence these pathways, it is concluded that Ca2+ exerts its primary effect by slowing passive diffusion. In support of this, Ca2+ also slowed 22Na+ efflux. In addition, ethanol-induced leakage of 86Rb+ was reversed by extracellular Ca2+, suggestive of a Ca2+-membrane phospholipid interaction.  相似文献   

9.
Elevated levels of intracellular Ca2+ activate a K+-selective permeability in the membrane of human erythrocytes. Currents through single channels were analysed in excised inside-out membrane patches. The effects of several ions that are known to inhibit K+ fluxes are described with respect to the single-channel events. The results suggest that the blocking ions can partly move into the channels (but cannot penetrate) and interact with other ions inside the pore. The reduction of single-channel conductance by Cs+, tetraethylammonium and Ba2+ and of single-channel activity by quinine and Ba2+ is referred to different rates of access to the channel. The concentration- and voltage-dependent inhibition by ions with measurable permeability (Na+ and Rb+) can be explained by their lower permeability, with single-file movement and ionic interactions inside the pore.  相似文献   

10.
The suppression of the cyclic nucleotide‐gated channel (CNGC) AtCNGC10 alters K+ transport in Arabidopsis plants. Other CNGCs have been shown to transport Ca2+, K+, Li+, Cs+ and Rb+ across the plasma membrane when expressed in heterologous systems; however, the ability of the AtCNGC10 channel to transport nutrients other than K+ in plants has not been previously tested. The ion fluxes along different zones of the seedling roots, as estimated by the non‐invasive ion‐specific microelectrode technique, were significantly different in two AtCNGC10 antisense lines (A2 and A3) in comparison to the wild type (WT). Most notably, the influxes of H+, Ca2+ and Mg2+ in the meristem and distal elongation zones of the antisense A2 and A3 lines were significantly lower than in the WT. The lower Ca2+ influx from the external media corresponded to a lower intracellular Ca2+ activity, which was estimated by fluorescence lifetime imaging measurements (FLIM). On the other hand, the intracellular pH values in the meristem zone of the roots of A2 and A3 seedlings were significantly lower (more acidic) than that of the WT, which might indicate a feedback block of H+ influx into meristematic cells caused by low intracellular pH. Under the control conditions, mature plants from the A2 and A3 lines contained significantly higher K+ and lower Ca2+ and Mg2+ content in the shoots, indicating disturbed long‐distance ion transport of these cations, possibly because of changes in xylem loading/retrieval and/or phloem loading. Exposing the plants in the flowering stage to various K+, Ca2+ and Mg2+ concentrations in the solution led to altered K+, Ca2+ and Mg2+ content in the shoots of A2 and A3 plants in comparison with the WT, suggesting a primary role of AtCNGC10 in Ca2+ (and probably Mg2+) transport in plants, which in turn regulates K+ transporters' activities.  相似文献   

11.
Abstract: The effects of four K+-channel inhibitors on synaptosomal free Ca2+ concentrations and 86Rb+ fluxes are analysed. 4-Aminopyridine, α-dendrotoxin, charybdotoxin, and tetraethylammonium all increase the free Ca2+ concentration, although their potencies differ widely. In each case, the elevation in free Ca2+ concentration is reversed by the subsequent addition of tetrodotoxin. The transient 86Rb+ efflux from preequilibrated synaptosomes induced with high concentrations of veratridine is partially inhibited by 4-aminopyridine and α-dendrotoxin. In contrast, when 4-aminopyridine or α-dendrotoxin is added to polarized synaptosomes, an enhanced86Rb+ flux is seen, both for uptake and for efflux with no change in the total 86Rb+/K+ content of the synaptosomes and with only a slight time-averaged plasma membrane depolarization (6.4 and 3.3 mV, respectively). The enhancements of flux by 4-aminopyridine or α-dendrotoxin are sensitive to ouabain and/or to tetrodotoxin. Furthermore, these flux changes show the same concentration dependencies as the blocked component of veratridine-stimulated 86Rb+ efflux, the elevation of free Ca2+ concentration, and the facilitation of glutamate exocytosis that are elicited by 4-aminopyridine or α-dendrotoxin. It is concluded that these findings support the proposal of spontaneous, repetitive firing of synaptosomes evoked by K+-channel inhibitors and that the enhanced 86Rb+ flux is a consequence of the activity of 4-aminopyridine- and α-dendrotoxin-insensitive K+ channels during these action potentials.  相似文献   

12.
The release of H+ during the oxalate-supported Ca2+ uptake in sarcoplasmic reticulum vesicles is kinetically coincident with the initial phase of Ca2+ accumulation. The Ca2+ uptake is increased and the H+ release is decreased in the presence of KCl and other monovalent chloride salts as expected for a H+-monovalent cation exchange. The functioning of the Ca2+-pump is disturbed by the presence of potassium gluconate and, to a lesser extent, of choline chloride. These salts do not inhibit the ATPase activity of Ca2+-permeable vesicles, suggesting a charge imbalance inhibition which is specially relevant in the case of gluconate. Therefore, K+, and also Cl, appear to be involved in secondary fluxes during the active accumulation of Ca2+. The microsomal preparation seems homogeneous with respect to the K+-channel, showing an apparent rate constant for K+ release of approximately 25 s–1 measured with the aid of86Rb+ tracer under equilibrium conditions. A Rb+ efflux, sensitive to Ca2+-ionophore, can be also detected during the active accumulation of Ca2+. The experimental data suggest that both monovalent cations and anions are involved in a charge compensation during the Ca2+ uptake and H+ release. Fluxes of these highly permeable ions would contribute to cancel the formation of a resting membrane potential through the sarcoplasmic reticulum membrane.  相似文献   

13.
Summary Bovine aortic endothelial cells (BAECs) respond to bradykinin with an increase in cytosolic-free Ca2+ concentration, [Ca2+] i , accompanied by an increase in surface membrane K+ permeability. In this study, electrophysiological measurement of K+ current was combined with86Rb+ efflux measurements to characterize the K+ flux pathway in BAECs. Bradykinin- and Ca2+-activated K+ currents were identified and shown to be blocked by the alkylammonium compound, tetrabutylammonium chloride and by the scorpion toxin,noxiustoxin, but not by apamin or tetraethylammonium chloride. Whole-cell and single-channel current analysis suggest that the threshold for Ca2+ activation is in the range of 10 to 100nm [Ca2+] i . The whole-cell current measurement show voltage sensitivity only at the membrane potentials more positive than 0 mV where significant current decay occurs during a sustained depolarizing pulse. Another K+ current present in control conditions, an inwardly rectifying K+ current, was blocked by Ba2+ and was not affected bynoxiustoxin or tetrabutylammonium chloride. Efflux of86Rb from BAEC monolayers was stimulated by both bradykinin and ionomycin. Stimulated efflux was blocked by tetrabutyl- and tetrapentyl-ammonium chloride and bynoxiustoxin, but not by apamin or furosemide. Thus,86Rb+ efflux stimulated by bradykinin and ionomycin has the same pharmacological sensitivity as the bradykinin- and Ca2+-activated membrane currents. The results confirm that bradykinin-stimulated86Rb+ efflux occurs via Ca2+-activated K+ channels. The blocking agents identified may provide a means for interpreting the role of the Ca2+-activated K+ current in the response of BAECs to bradykinin.  相似文献   

14.
Summary This study concerns the properties of rapid K+ and Cl transport pathways that are present in the (H++K+)-ATPase membrane from stimulated, and secreting, gastric oxyntic cells. Ion permeabilities in the isolated stimulation-associated vesicles were monitored via the rates of H+ efflux under conditions of exclusive H+/K+ counterflux or H+–Cl co-efflux, as well as by comparison of equilibration rates for86Rb and36Cl under conditions of equilibrium exchange and unidirectional salt flux. These latter studies suggest that Rb+ and Cl pathways are conductive and independent. In spite of the functional independence of the ion pathways, several divalent cations inhibit Rb+ and Cl isotopic exchange as well as the H+ efflux that is dependent on either K+ or anion (Cl, SCN, NO2) fluxes. Zn2+ is the more potent inhibitor, reducing by 50% the sensitive component of K+, Cl, and NO2 fluxes at about 20 m; Mn2+ has a similar effect at 200 m. Ni2+ and Co2+ were roughly equipotent to Mn2+ while Mg2+ and Ca2+ had not inhibitory effect. These results suggest that the stimulation-induced permeabilities, while functioning independently, may be physically linked, i.e., residing within a single entity. In similar studies carried out in (H++K+)-ATPase vesicles obtained from nonstimulated cells, no vestiges of sensitivity to the inhibitory divalent cations could be detected. The implications of these findings for the physiology of the oxyntic cell in the context of a model for membrane fusion are discussed.  相似文献   

15.
Ice crystal formation temperature was determined in the region of the crown in one group of 7-day-old intact unhardened high-salt plants of winter wheat (Triticum aestivum L. cv. Weibulls Starke II) with TA (Thermal Analysis) and DTA (Differential Thermal Analysis) methods. After exposure of another group of plants, grown for the first 7 days in the same way as the first group, to various sub-zero temperatures (-1 to 5°C), influx in roots of Rb+(86Rb+) and Ca2+(45Ca2+) and contents of K+ and Ca2+ were determined at intervals during 7 days of recovery. Ice crystal formation in the crown tissue was probably extracellular and took place at about -4°C. There was a large loss of K+ from the roots after treatment at sub-zero temperatures. This loss increased as the temperature of the sub-zero treatment decreased. During recovery, roots of plants exposed to -1, -2 and -3°C gradually reabsorbed K+. Reabsorption of K+ in roots of plants exposed to -4°C was greatly impaired. Rb+ influx decreased and Ca2+ influx increased after sub-zero temperature treatments of the plants. Active Rb+ influx mechanisms and active extrusion of Ca2+ were impaired or irreversibly damaged by the exposure. While Rb+ influx mechanisms were apparently repaired during recovery in plants exposed to temperatures down to -3°C, Ca2+ extrusion mechanisms were not. The temperature for ice crystal formation in the region of the crown tissue coincides with the temperature at which the plants lost the ability to reabsorb K+ and to repair Rb+ influx mechanisms during the recovery period. Plants were lethally damaged at temperatures below ?4°C.  相似文献   

16.
Influx of Rb+(86Rb+) and Ca2+ (45Ca2+) in roots of intact winter wheat (Triticum aestivum L. cv. Weibulls Starke II) was determined at intervals before, during and after exposure to cold acclimation conditions (2°C and 8 h light period). The plants were grown in nutrient medium of two ionic strengths. During the initial two weeks of growth at 16°C and 16 h light period, Rb+ influx into roots decreased with increasing age, probably as a consequence of a decreasing proportion of metabolically active roots. The presence of 10?4M 2,4-dinitrophenol (DNP) reduced Rb+ influx to a low and constant level, indicating that metabolic influx was the dominant process. In contrast, Ca2+ influx in plants grown in full strength nutrient solution was higher in the presence than in the absence of DNP. This effect may have been due to an active extrusion mechanism mediating re-export of absorbed Ca2+(45Ca2+) during the uptake experiment. With the metabolic uncoupler inhibiting such extrusion the Ca2+(45Ca2+) influx mesured would increase. During cold treatment, Rb+ influx remained at a low level, and was further decreased when DNP was present in the uptake solution. This effect may have been due to inhibition of residual active influx of Rb+ at 2°C by the uncoupler and/or to a decrease in membrane permeability. In contrast to Rb+, Ca2+ influx increased during cold treatment, which could again be explained as inhibition of re-export. The presence of DNP reduced Ca2+ influx at 2°C, indicating decreased membrane permeability by DNP at low temperature. After transfer of plants from cold acclimation conditions to 16°C, Rb+ and Ca2+ influx increased in plants grown at both ionic strengths. Influx levels were independent of the length of the cold acclimation period (1, 6 and 8 weeks), but the patterns were different for the two ions. After each of the cold acclimation periods, Rb+ influx increased during the first week and decreased or remained at the same level during the second week, while Ca2+ influx always decreased during the second week of post-cold treatment.  相似文献   

17.
The Ussing chamber technique was used to measure unidirectional Rb+ fluxes under short-circuit conditions across tissue sheets from proximal, central, and distal jejunum of rats. Whereas the proximal and central parts of the jejunum did not show any net transport of Rb+, there was a net secretion of around 0.2 μmol hr−1 cm−2 in the distal segment. This secretion could not be influenced significantly by mucosal application of K+ channel blockers such as Ba2+ (5 mm), tetraethylammonium (20 mm) or quinine (1 mm). Serosal ouabain (1 mm) blocked net secretion by increasing mucoserosal flux. Blockers of H+/K+ ATPases could not alter net fluxes of Rb+. Stimulation of Cl secretion by forskolin (10 μm) or of Na+ absorption by serine (10 mm) failed to influence the observed secretion of Rb+. Adrenaline (10 μm) also had no effect on Rb+ fluxes. Blocking Na+/H+ exchange by 5-(N-Ethyl-N-isopropyl)-amilorid (100 μm) blocked net secretion by increasing mucoserosal flux, as did the addition of Na+ acetate (30 mm) to the mucosal solution. We conclude that the distal jejunum of the rat secretes K+ under short-circuit conditions. This secretion does not seem to occur via K+ channels, but through a pH dependent mechanism. Received: 16 February 1999/Revised: 29 June 1999  相似文献   

18.
K+ channels, membrane voltage, and intracellular free Ca2+ are involved in regulating proliferation in a human melanoma cell line (SK MEL 28). Using patch-clamp techniques, we found an inwardly rectifying K+ channel and a calcium-activated K+ channel. The inwardly rectifying K+ channel was calcium independent, insensitive to charybdotoxin, and carried the major part of the whole-cell current. The K+ channel blockers quinidine, tetraethylammonium chloride and Ba2+ and elevated extracellular K+ caused a dose-dependent membrane depolarization. This depolarization was correlated to an inhibition of cell proliferation. Charybdotoxin affected neither membrane voltage nor proliferation. Basic fibroblast growth factor and fetal calf serum induced a transient peak in intracellular Ca2+ followed by a long-lasting Ca2+ influx. Depolarization by voltage clamp decreased and hyperpolarization increased intracellular Ca2+, illustrating a transmembrane flux of Ca2+ following its electrochemical gradient. We conclude that K+ channel blockers inhibit cell-cycle progression by membrane depolarization. This in turn reduces the driving force for the influx of Ca2+, a messenger in the mitogenic signal cascade of human melanoma cells. Received: 9 May 1995/Revised: 30 January 1996  相似文献   

19.
Influx of Rb+(86Rb+) and Ca2+(45Ca2+) was determined in roots of winter wheat (Triticum aestivum L. cv. Weibulls Starke II) after 14 days at 16°C/16 h light, after 1 and 8 weeks of cold acclimation (2°C/8 h light) and at intervals after deacclimation (16°C/16 h light) for up to 14 days. The plants were cultivated at 3 ionic strengths: 100, 10 and 1% of a full strength nutrient solution, containing 3.0 mM K+ and 1.0 mM Ca2+. K+ concentrations in roots and shoots increased during cold treatment, while Ca2+ in the roots decreased. In the shoots Ca2+ concentrations remained the same. Influx of Rb+ as a function of average K+ concentration in the roots of 14-day-old, non-cold-treated plants was high at a certain K+ level in the root and decreased at higher root K+ levels (negative feedback). The pattern for Ca2+ influx versus average concentration of Ca2+ in the root was the reverse. Independent of duration of treatment (1–8 weeks), cold acclimation partly changed the regulation of Rb+ influx, so that it became less dependent upon negative feedback and more dependent on the ionic strength of the cultivation solution. After exposure to 2°C, Ca2+ influx increased at high Ca2+ concentrations in the root as compared with influx in roots of 14-day-old non-cold-treated plants. Under deacclimation, Ca2+ influx gradually decreased again, and reached the level observed before cold treatment within 7–14 days at 16°C; the number of days depending on the exposure time at 2°C. It is suggested that Rb+(K+) influx became adjusted to low temperature and that abscisic acid (ABA) may be involved in this mechanism. It is also suggested that extrusion of Ca2+ was impaired and/or Ca2+ channels were activated at 2°C in roots of plants grown in the full-strength solution and that extrusion was gradually restored and/or Ca2+ channels were closed under deacclimation conditions.  相似文献   

20.
《Cell calcium》2016,59(6):577-588
Rises in cytosolic Ca2+ concentration ([Ca2+]cyt) are central in platelet activation, yet many aspects of the underlying mechanisms are poorly understood. Most studies examine how experimental manipulations affect agonist-evoked rises in [Ca2+]cyt, but these only monitor the net effect of manipulations on the processes controlling [Ca2+]cyt (Ca2+ buffering, sequestration, release, entry and removal), and cannot resolve the source of the Ca2+ or the transporters or channels affected. To investigate the effects of protein kinase C (PKC) on platelet Ca2+ signalling, we here monitor Ca2+ flux around the platelet by measuring net Ca2+ fluxes to or from the extracellular space and the intracellular Ca2+ stores, which act as the major sources and sinks for Ca2+ influx into and efflux from the cytosol, as well as monitoring the cytosolic Na+ concentration ([Na+]cyt), which influences platelet Ca2+ fluxes via Na+/Ca2+ exchange. The intracellular store Ca2+ concentration ([Ca2+]st) was monitored using Fluo-5N, the extracellular Ca2+ concentration ([Ca2+]ext) was monitored using Fluo-4 whilst [Ca2+]cyt and [Na+]cyt were monitored using Fura-2 and SFBI, respectively. PKC inhibition using Ro-31-8220 or bisindolylmaleimide I potentiated ADP- and thrombin-evoked rises in [Ca2+]cyt in the absence of extracellular Ca2+. PKC inhibition potentiated ADP-evoked but reduced thrombin-evoked intracellular Ca2+ release and Ca2+ removal into the extracellular medium. SERCA inhibition using thapsigargin and 2,5-di(tert-butyl) l,4-benzohydroquinone abolished the effect of PKC inhibitors on ADP-evoked changes in [Ca2+]cyt but only reduced the effect on thrombin-evoked responses. Thrombin evokes substantial rises in [Na+]cyt which would be expected to reduce Ca2+ removal via the Na+/Ca2+ exchanger (NCX). Thrombin-evoked rises in [Na+]cyt were potentiated by PKC inhibition, an effect which was not due to altered changes in non-selective cation permeability of the plasma membrane as assessed by Mn2+ quench of Fura-2 fluorescence. PKC inhibition was without effect on thrombin-evoked rises in [Ca2+]cyt following SERCA inhibition and either removal of extracellular Na+ or inhibition of Na+/K+-ATPase activity by removal of extracellular K+ or treatment with digoxin. These data suggest that PKC limits ADP-evoked rises in [Ca2+]cyt by acceleration of SERCA activity, whilst rises in [Ca2+]cyt evoked by the stronger platelet activator thrombin are limited by PKC through acceleration of both SERCA and Na+/K+-ATPase activity, with the latter limiting the effect of thrombin on rises in [Na+]cyt and so forward mode NCX activity. The use of selective PKC inhibitors indicated that conventional and not novel PKC isoforms are responsible for the inhibition of agonist-evoked Ca2+ signalling.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号