共查询到19条相似文献,搜索用时 546 毫秒
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采用离子选择电极测量羊浦肯野纤维细胞膜内钠离子活度(~(ai)N_a),细胞间钾离子活度(a~ok)及细胞膜电位(v_m),观察不同浓度低钠,无钙液对其影响,在无钙低钠液中,细胞内Na~+逐出,α~iNa 降低,其变化速率,幅值与[Na]_o 相关,同时也受细胞 a~iNa 初始水平(aiNa(o))的影响。aiNa 下降6min 时的稳态水平与[Na]_o 呈直线正相关,这些结果表明,[Na]_o 降低时,细胞膜钠泵活动加强,细胞内 Na~+逐出增加,其最终结果是使 Na+跨膜梯度维持相对稳定,因而可以认为是 Na~+跨膜梯度而不是单纯的细胞内 Na~+控制膜钠泵活动。在低 Na~+液引起细胞内 Na~+主动逐出增加的同时,细胞膜出现超极化,[Na]_o 愈低,膜超极化程度愈高,从低钠液引起的 a~i_(Na),V_m,α~o_k 变化之间的时程关系看,膜超极化主要由加大的外向泵电流引起,同时发生的细胞间 K~+浓度变化对其也有一定影响。 相似文献
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Activation of Na+ /H+ exchange on rat preadipocyte plasma membrane and its role in cell proliferation and differentiation 总被引:1,自引:0,他引:1
An in vitro cultured rat perirenal preadipocyte (PA) was established as a model system to investigate the role of the intracellular pH (pHi) and of the Na~ /H~ exchanger during PA proliferation and differentiation, pH sensitive probe, 2' ,7'-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein(BCECF), was employed to measure the pHi of PA and to determine the Na~ /H~ exchange activity. The results showed that there was Na~ /H~ exchange activity in the plasma membrane of PA, FCS stimulated DNA synthesis measured by ~3H-TdR incorporation, and the activation of Na~ /H~ exchanger resulted in phi increase (nearly 0.2 pH unit) within 2 min. Ethyl-isopropyl-amiloride (EIPA), a specific Na~ /H~ exchange inhibitor, inhibited Na~ /H~ exchange activity and DNA synthesis. In the absence of serum insulin did not stimulate DNA synthesis but did induce PA differentiation characterized by the appearance of adiposome in the cell and the enhancement of glycerol-3-phosphate dehydrogenase (G_3PDHase) activity. Meantime, insu 相似文献
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黑河天涝池流域典型林分生态水文化学特征 总被引:2,自引:0,他引:2
采集了黑河天涝池流域典型林分林外雨、穿透雨、树干径流和枯透水,并检测水体pH值和12种离子(K~+、Ca~(2+)、Na~+、Mg~(2+)、NH_4~+、Cu~(2+)、Zn~(2+)、Pb~(2+)、Cd~(2+)、Cl~-、SO_4~(2-)、NO_3~3)的质量浓度。结果表明:天涝池流域大气降水pH均值为7.74,呈碱性,降水中离子绝对质量浓度较低,最高的是NO_3~-,质量浓度为1.1111 mg/L,最低的为Na~+,质量浓度为0.0108 mg/L;两种林分冠层有降低降雨pH值的作用,青海云杉林冠层对NH_4~+有升高作用,祁连圆柏林冠层对NH_4~+有降低作用,两种林冠层对NO_3~-和Cu~(2+)质量浓度有降低作用,对其它离子质量浓度均表现为升高作用;两种林分树干径流有提高穿透雨pH值的作用,与穿透雨相比,两种林分树干径流中阴离子均有升高,圆柏树干径流中所有阳离子质量浓度均有下降,云杉树干径流中Ca~(2+)、K~+、Mg~(2+)和Na~+减少,NH_4~+和Cu~(2+)增加;典型林分枯透水有提升穿透雨pH值的作用,与穿透雨相比,两种林分枯透水中阴离子质量浓度均有升高,云杉枯透水各阳离子均有降低,圆柏枯透水中Ca~(2+)、K~+和Mg~(2+)质量浓度升高,NH_4~+、Na~+和Cu~(2+)质量浓度下降;在采集的所有样本中,Pb~(2+)和Cd~(2+)均未检出,而Zn~(2+)仅在云杉树干径流中检出。 相似文献
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符云峰 《中国生物化学与分子生物学报》1988,4(1):84-87
本研究确定了在0℃条件下,(Na~++K~+)-ATP酶纯化制备物与5mmol/L Na~+或Mg~(2+)在5mmol/L咪唑(pH7.4)环境中预保温30分钟,然后进行磷酸化,可以获得最高磷酸化水平,Na~+或Mg~(2+)的K_(0.5)值分别为0.29mmol/L或0.35mmol/L;以ADP代替Na~+和Mg~(2+)与酶预保温,对E_2向E_1转变无任何影响,而与Na~+、Mg~(2+)一起存在时则能加强Na~+及Mg~2的预保温效果。 相似文献
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《生态学杂志》2017,(3)
运用相关分析、因子分析和富集因子等对布哈河流域2014年丰水期河水样品主要离子浓度特征及来源进行了分析。结果表明:布哈河流域河水p H值的变化范围为7.91~9.21,平均值为8.43;EC的变化范围为133~871μS·cm~(-1),平均值为383μS·cm~(-1);河水中SO_4~(2-)、Ca~(2+)、Cl-、Na~+、 Mg~(2+)、NO3-、K+和NH_4~+的平均离子浓度分别为60.2、33.1、31.1、14.8、12.8、8.1、1.5和0.3μeq·L-1;阳离子浓度的大小顺序为Ca~(2+)Na~+ Mg~(2+)K+NH_4~+,而主要阴离子浓度的大小顺序SO_4~(2-)Cl-NO_3~-; Mg~(2+)、Ca~(2+)和SO_4~(2-)的离子浓度随着海拔的增加呈增加的趋势,Na+、Cl-和NO_3~-随着海拔的增加表现出减小的趋势,但K~+和NH_4~+不随海拔梯度的改变而变化;布哈河全流域的水化学类型为SO_4~(2-)-Ca~(2+)-Cl-- Mg~(2+);布哈河流域的水化学主要受控于蒸发岩和碳酸盐岩风化产物的影响。 相似文献
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本研究采用羊心脏浦肯野纤维标本,按其静息膜电位水平分为两组:;≥-75mV的为正常组,-40~-55mV为部分除极组。在降低细胞内外Na~ 跨膜梯度的情况下,用细胞内离子选择电极监测细胞内Na~ 主动逐出速率以测量细胞膜钠泵活动。结果表明,除极组细胞内Na~ 活度(a~1Na)对照值及在预先提高细胞内Na~ 水平的情况下所测得的Na~ 逐出速半与正常组无显著差别。因而认为,自然部分除极的浦肯野纤维膜钠泵机能并无不可逆转的损伤。其除极原因可能主要与膜对K~ 的通透性降低有关。 相似文献
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《生物化学与生物物理进展》2017,(7)
正Dear editor,Light-driven Na~+ pumping rhodopsins(Na Rs),the next generation optogenetic tools,are a family of seven-helical transmembrane proteins and covalently link to a retinal chromophore~([1]).Na Rs have a putative Na~+ -conducting motif of Asn112-Asp116-Gln123~([1]),homologous to the H~+-conducting motif of Asp85-Thr89-Asp96 in H~+ pump bacteriorhodopsin(BR)~([2]). 相似文献
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《植物生态学报》2017,(4)
为了进一步从离子动态运输方面了解沙枣(Elaeagnus angustifolia)耐盐机制和揭示沙枣种源间的K~+/Na~+平衡调控差异,该研究利用非损伤微测技术(non-invasive micro-test technology,NMT)测定银川种源(盐敏感型)和阿拉尔种源(耐盐型)沙枣幼苗根系在3种不同NaCl处理方式下的离子流:1)在150 mmol·L–1 NaCl胁迫24 h后的Na~+和K~+离子流;2)NaCl瞬时处理后的K~+和H~+的动态离子流;3)先NaCl胁迫24 h,再用Na~+/H~+逆向转运体抑制剂阿米洛利(Amiloride)和K~+通道抑制剂氯化四乙胺(TEA)处理后的Na~+和K~+离子流。结果表明:NaCl胁迫24 h后,沙枣根系Na~+和K~+外排净流量显著增加,并且银川种源沙枣幼苗根系Na~+净流量显著低于阿拉尔种源,净流量分别为720和912 pmol·cm~(–2)·s~(–1),而K~+外流净流量显著高于阿拉尔种源。瞬时NaCl处理后,沙枣根系K~+的外流迅速增加,并且银川种源的K~+外排净流量始终高于阿拉尔种源,而H~+由内流转为外排,阿拉尔种源的H~+净外流量大于银川种源。NaCl和NaCl+Amiloride处理下,阿拉尔种源沙枣幼苗Na~+外流的净流量均大于银川种源,但K~+外流的净流量均小于银川种源,而在对照和NaCl+TEA处理下,Na~+和K~+的净流量在两个种源间无明显差异。研究证明NaCl胁迫造成根系Na~+积累和K~+外流,沙枣幼苗为减少Na~+积累,通过根系Na~+/H~+逆向转运体将Na~+从体内排出,并且耐盐型种源沙枣幼苗根系在NaCl胁迫时能更好地维持体内的K~+/Na~+平衡,其原因主要在于具有较强的Na~+外排能力和较弱的K~+流失。该研究可以为进一步发掘优良耐盐沙枣种质资源提供理论参考依据。 相似文献
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To study the role of intracellular pH (pHi) in catecholamine secretion and the regulation of pHi in bovine chromaffin cells, the pH-sensitive fluorescent indicator [2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein] was used to monitor the on-line changes in pHi. The pHi of chromaffin cells at resting state is approximately 7.2. The pHi was manipulated first by incubation of the cells with NH4+, and then the solution was replaced with a NH4(+)-free solution to induce acidification of the cytoplasm. The pHi returned toward the basal pH value after acidification within 5-10 min in the presence of Na+ or Li+, but the pHi stayed acidic when Na(+)-free buffers were used or in the presence of amiloride and its analogues. These results suggest that the pH recovery process after an acid load is due to the Na+/H+ exchange activity in the plasma membrane of the chromaffin cells. The catecholamine secretion evoked by carbachol and Na+ removal was enhanced after the cytoplasm had been made more acidic. It appears that acidic pH favors the occurrence of exocytosis. 相似文献
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Nobuyuki Yanagihara Kenji Yokota Hideyuki Kobayashi Akihiko Wada Yasuhito Uezono Futoshi Izumi 《Journal of neurochemistry》1990,54(5):1626-1631
We investigated the presence of Na+/H+ exchange in cultured bovine adrenal medullary cells. The intracellular pH in control cells measured by 5,5-dimethyl[2-14C]oxazolidine-2,4-dione was 7.13 +/- 0.02 (n = 6). Removal of Na+ from the incubation medium shifted the intracellular pH down to 6.67 +/- 0.12 (n = 6). Reintroduction of Na+ to the medium caused a rapid recovery in intracellular pH to 7.20-7.30 that was associated with an increase in uptake of 22Na+ by the cells. Both increases in intracellular pH and uptake of 22Na+ were inhibited by amiloride, an inhibitor of Na+/H+ exchange. The recovery of intracellular pH by addition of Na+ was partially inhibited by quinidine, another inhibitor of Na+/H+ exchange, but not by 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid, an anion-exchange (Cl-/HCO3-) inhibitor. Li+ could substitute for Na+ in the recovery of intracellular pH. Carbachol caused an increase in intracellular pH from 7.12 +/- 0.01 to 7.21 +/- 0.02 (n = 10). This increase in intracellular pH caused by carbachol was inhibited by amiloride. These results suggest the existence of an amiloride-sensitive Na+/H+ exchange that regulates the intracellular pH in adrenal medullary cells. 相似文献
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The aim of this study was to investigate the effect of NH4+ on the intracellular pH in TALH SVE.1 cells derived from the medullary thick ascending limb of Henle's loop (TALH) of rabbit kidney. These cells are specialized to perform NH4+ transport in vivo. Intracellular pH was monitored by 31P-NMR. The steady state intracellular pH (pHi) under standard conditions was 7.24 +/- 0.04 (n = 46). Exposure to NH4Cl resulted in an initial intracellular acidification of the TALH SVE.1 cells, followed by a recovery to the initial steady-state pHi value. The NH4(+)-induced acidification followed saturation kinetics up to 20 mM NH4Cl (delta pHmax = 0.2 pHunits). Half-maximal acidification was observed at 0.6 mmol/l. The intracellular acidification due to NH4Cl exposure was completely inhibited by 0.1 mM of the diuretic bumetanide, an inhibitor of the Na+/K+/2Cl- cotransporter. The effect of bumetanide was dose-dependent and a Ki value of 8.10(-7) M was calculated. NH4+ influx via K+ channels or the (Na+ + K+)ATPase could not be detected. pHi recovery to the initial value was caused mainly by amiloride-sensitive Na+/H+ exchange and to a lesser extent by an amiloride-insensitive system, which was not studied in detail. In the presence of bumetanide, pulses of high concentrations of NH4Cl induced small intracellular alkalinizations. From these experiments, an intrinsic buffer capacity (beta i) in TALH SVE.1 cells of 26 +/- 3 mM x pH-1 (pHi = 7.65) was determined. It could also be shown that the TALH SVE.1 cells exhibit maximal 'functional buffer capability' between pHout 6.9 and 7.3. Within these limits the cells can maintain their intracellular pH at a constant level, even though the extracellular pH changes. These data strongly suggest that the Na+/K+/2Cl- cotransporter is the main site of NH4+ entry into rabbit thick ascending limb cells in culture. A high intracellular buffer capacity and potent acid extrusion mechanism cooperate in counteracting the intracellular acidification caused by NH4+ influx into the cell. 相似文献
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Guo H. Zhang Edward J. Cragoe Jr. James E. Melvin 《The Journal of membrane biology》1992,129(3):311-321
The regulation of intracellular pH (pHi) in rat sublingual mucous acini was monitored using dual-wavelength microfluorometry of the pH-sensitive dye BCECF (2',7'-biscarboxyethyl-5(6)-carboxyfluorescein). Acini attached to coverslips and continuously superfused with HCO3(-)-containing medium (25 mM NaHCO3/5% CO2; pH 7.4) have a steady-state pHi of 7.25 +/- 0.02. Acid loading of acinar cells using the NH4+/NH3 prepulse technique resulted in a Na(+)-dependent, MIBA-inhibitable (5-(N-methyl-N-isobutyl) amiloride, Ki approximately 0.42 microM) pHi recovery, the kinetics of which were not influenced by the absence of extracellular Cl-. The rate and magnitude of the pHi recovery were dependent on the extracellular Na+ concentration, indicating that Na+/H+ exchange plays a critical role in maintaining pHi above the pH predicted for electrochemical equilibrium. When the NH4+/NH3 concentration was varied, the rate of pHi recovery was enhanced as the extent of the intracellular acidification increased, demonstrating that the activity of the Na+/H+ exchanger is regulated by the concentration of intracellular protons. Switching BCECF-loaded acini to a Cl(-)-free medium did not significantly alter resting pHi, suggesting the absence of Cl-/HCO3- exchange activity. Muscarinic stimulation resulted in a rapid and sustained cytosolic acidification (t 1/2 < 30 sec; 0.16 +/- 0.02 pH unit), the magnitude of which was amplified greater than two-fold in the presence of MIBA (0.37 +/- 0.05 pH unit) or in the absence of extracellular Na+ (0.34 +/- 0.03 pH unit). The agonist-induced intracellular acidification was blunted in HCO3(-)-free media and was inhibited by DPC (diphenylamine-2-carboxylate), an anion channel blocker. In contrast, the acidification was not influenced by removal of extracellular Cl-. The Ca2+ ionophore, ionomycin, mimicked the effects of stimulation, whereas preloading acini with BAPTA (bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetra-acetic acid) to chelate intracellular Ca2+ blocked the agonist-induced cytoplasmic acidification. The above results indicate that during muscarinic stimulation an intracellular acidification occurs which: (i) is partially buffered by increased Na+/H+ exchange activity; (ii) is most likely mediated by HCO3- efflux via an anion channel; and (iii) requires an increase in cytosolic free [Ca2+]. 相似文献
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Continuous intracellular pH (pHi) measurements were performed in SIRC rabbit corneal epithelial cells using the pH-sensitive absorbance of intracellularly trapped 5(and 6)-carboxy-4',5'-dimethylfluorescein. Steady-state pHi in nominally bicarbonate free Ringer's solution averaged 6.87 +/- 0.02 (mean +/- S.E., n = 53). After intracellular acidification induced by the NH4Cl-prepulse technique, there was a sodium-dependent pHi recovery towards the normal steady-state pHi. The initial pHi recovery rate was a saturable function of extracellular sodium concentration with an apparent Km for external sodium of about 25 mM and a Vmax of about 0.28 pH units/min. Virtually no pHi recovery was observed in the absence of extracellular sodium. Sodium removal during steady state acidified the cells by 0.36 +/- 0.05 pH units (mean +/- S.E., n = 13) within 5 min. There was a dose-dependent inhibition of pHi recovery after NH4Cl prepulse by amiloride with an IC50 of about 15 microM. Amiloride in a concentration of 1 mM almost completely abolished pHi recovery. Amiloride (1 mM) applied during steady state induced an intracellular acidification of 0.2 +/- 0.03 pH units (mean +/- S.E., n = 7) within 5 min. These findings suggest that a Na+/H+ exchange is present in SIRC rabbit corneal epithelial cells. Na+/H+ exchange seems to be the major process involved in pHi recovery in SIRC cells after an intracellular acid load. Na+/H+ exchange also plays a role in the maintenance of steady-state pHi. 相似文献
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Sealed membrane vesicles of Acholeplasma laidlawii were obtained by controlled lysis of carotenoid-rich intact cells. An imposed delta pH was created by loading membrane vesicles or intact Acholeplasma laidlawii cells with 0.25 M NH4Cl and diluting them into 0.25 M choline chloride. The passive efflux of NH3 from the membrane vesicles or cells resulted in the creation of a delta pH (inside acid) that could be visualized by the quenching of the fluorescence of the weak base acridine orange. Whereas with isolated membrane vesicles, the fluorescence was dequenched by the addition of Na+, with intact cells, K+ in addition to Na+ was required. These results strongly suggest a Na+/H+ exchange activity that in intact Acholeplasma laidlawii cells is K+-dependent. The possible role of the Na+/H+ exchange activity in pH homeostasis at the more alkaline pH range, as well as in the extrusion of excess Na+ from the cells is discussed. 相似文献
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Apical membrane H+ extrusion in the renal outer medullary collecting duct, inner stripe, is mediated by a Na(+)-independent H+ pump. To examine the regulation of this transporter, cell pH and cell Ca2+ were measured microfluorometrically in in vitro perfused tubules using 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein and fura-2, respectively. Apical membrane H+ pump activity, assayed as cell pH recovery from a series of acid loads (NH3/NH+4 prepulse) in the total absence of ambient Na+, initially occurred at a slow rate (0.06 +/- 0.02 pH units/min), which was not sufficient to account for physiologic rates of H+ extrusion. Over 15-20 min after the initial acid load, the rate of Na(+)-independent cell pH recovery increased to 0.63 +/- 0.09 pH units/min, associated with a steady-state cell pH greater than the initial pre-acid load cell pH. This pattern suggested an initial suppression followed by a delayed activation of the apical membrane H+ pump. Replacement of peritubular Na+ with choline or N-methyl-D-glucosamine resulted in an initial spike increase in cell Ca2+ followed by a sustained increase in cell Ca2+. The initial rate of Na(+)-independent cell pH recovery could be increased by elimination of the Na+ removal-induced sustained cell Ca2+ elevation by: (a) performing studies in the presence of 135 mM peritubular Na+ (1 mM peritubular amiloride used to inhibit basolateral membrane Na+/H+ antiport); (b) clamping cell Ca2+ low with dimethyl-BAPTA, an intracellular Ca2+ chelating agent; or (c) removal of extracellular Ca2+. Cell acidification induced a spike increase in cell Ca2+. The late acceleration of Na(+)-independent cell pH recovery was independent of Na+ removal and of the method used to acidify the cell, but was eliminated by prevention of the cell Ca2+ spike and markedly delayed by the microfilament-disrupting agent, cytochalasin B. This study demonstrates that peritubular Na+ removal results in a sustained elevation in cell Ca2+, which inhibits the apical membrane H+ pump. In addition, rapid cell acidification associated with a spike increase in cell Ca2+ leads to a delayed activation of the H+ pump. Thus, cell Ca2+ per se, or a Ca(2+)-activated pathway, can modulate H+ pump activity. 相似文献