共查询到20条相似文献,搜索用时 531 毫秒
1.
YUKIO FUJINO KEIKO MITSUNAGA IKUO YASUMASU 《Development, growth & differentiation》1987,29(6):591-597
Embryos kept with omeprazole, a specific H+ , K+ -ATPase inhibitor, in a period of development between the mesenchyme blastula and the pluteus corresponding stage became abnormal plutei having quite small spicules, somewhat poor pluteus arms and apparently normal archenterons. In micro-mere-derived cells, kept with omeprazole at pH 8.2 in a period between 15 and 40 hr of culture at 20°C, omeprazole strongly inhibited spicule formation but did not block the outgrowth of pseudopodial cables, in which spicule rods were to be formed. These indicate that omeprazole probably exerts no obvious inhibitory effects other than spicule rods formation. Omeprazole-sensitive H+ , K+ -ATPase, an H+ pump, seems to be indispensable for CaCO3 deposition (formation of spicule rod) in these spicule forming cells. H+ , produced in overall reaction for CaCO3 formation: Ca2+ + CO2 +H2 O°CaCO3 +2H+ , is probably released from the cells by this H+ pump and hence, this reaction tends to go to CaCO3 production to form spicule rods. Omeprazole, known to become effective following its conversion to a specific inhibitor of H+ , K+ -ATPase at acidic pH, is able to inhibit formation of spicule rod at alkaline pH in sea water. This is probably due to an acidification of sea water near the cell surface by H+ ejection in H+ , K+ -ATPase reaction. 相似文献
2.
In embryos of the sea urchin, Hemicentrotus pulcherrimus , as well as in cultured cells derived from isolated micromeres, spicule formation was inhibited by allylisothiocyanate, an inhibitor of H+ , K+ -ATPase, at above 0.5 μM and was almost completely blocked at above 10 μM. Amiloride, an inhibitor of Na+ , H+ antiporter, at above 100 μM exerted only slight inhibitory effect, if any, on spicule formation. Intravesicular acidification, determined using [ dimethylamine -14 C]-aminopyrine as a pH probe, was observed in the presence of ATP and 200 mM KCl in microsome fraction obtained from embryos at the post gastrula stage, at which embryos underwent spicule calcification. Intravesicular acidification and K+ -dependent ATPase activity were almost completely inhibited by allylisothiocyanate at 10 μM. Allylisothiocyanate-sensitive ATPase activity was found mainly in the mesenchyme cells with spicules isolated from prisms. H+ , K+ -ATPase, an H+ pump, probably mediates H+ release to accelerate CaCO3 deposition from Ca2+ , CO2 and H2 O in the primary mesenchyme cells. Intravesicular acidification was stimulated by valinomycin at the late gastrula and the prism stages but not at the pluteus stage. K+ permeability probably increases after the prism stage to activate H+ release. 相似文献
3.
The effects of external K+ , H+ and Ca2+ concentrations on the intracellular K+ concentration, [K+ ]i, and the K+ -ATPase activity in 2-day-old mung bean roots [ Vigna mungo (L.) Hepper] were investigated. [K+ ]i, in mung bean roots was markedly decreased by external K+ or H+ stress and did not recover the initial value even after the stress was removed. This decrease in [K+ ]i, gradually disappeared with the addition of (Ca2+ . Ca2+ may offset the harmful effects of ion stress. Ca2+ seems to have two effects on K+ transport; control of K+ permeability and activation of K+ uptake, although K+ -ATPase activity was inhibited by Ca2+ concentrations higher than 10–4 M. We suggest that Ca2+ activates K+ uptake indirectly through the acidification of the cytoplasm. 相似文献
4.
Toshio Matsuda Yosuke Murata Koh-ichi Tanaka Rie Hosoi Masako Hayashi Kazuko Tamada Kazuhiro Takuma Akemichi Baba 《Journal of neurochemistry》1996,66(2):511-516
Abstract: We have previously reported that insulin/insulin-like growth factor (IGF)-I induced the α1 isoform of Na+ ,K+ -ATPase in cultured astrocytes. In this study the effects of insulin/IGF-I on Na+ ,K+ -ATPase activity and cell proliferation were examined in astrocytes cultured under the various conditions, to test the possible involvement of the enzyme activity in the mitogenic action of IGF-I on astrocytes. Insulin increased Na+ ,K+ -ATPase activity and stimulated cell proliferation in subconfluent astrocytes (cultured for 7–14 days in vitro). In contrast, these effects were not observed in confluent cells (cultured for 28 days). Furthermore, insulin stimulated neither the enzyme activity nor [3 H]thymidine incorporation in astrocytes preincubated in fetal calf serum-free medium for 2 days (quiescent cells) and treated with dibutyryl cyclic AMP (differentiated cells). The increases in Na+ ,K+ -ATPase activity and expression of the α1 mRNA preceded the mitogenic effect. 125 I-IGF-I binding experiment showed that all the cells used here had similar binding characteristics. The insulin-induced increase in enzyme activity was not affected by 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), and it was observed even in Ca2+ -free medium. The stimulation by IGF-I of [3 H]thymidine incorporation was attenuated by ouabain and a low external K+ level. These findings suggest that stimulation of Na+ ,K+ -ATPase activity is involved in the mitogenic action of IGF-I on cultured astrocytes. 相似文献
5.
Abstract: The excitatory amino acid glutamate was previously shown to stimulate aerobic glycolysis in astrocytes by a mechanism involving its uptake through an Na+ -dependent transporter. Evidence had been provided that Na+ ,K+ -ATPase might be involved in this process. We have now measured the activity of Na+ ,K+ -ATPase in cultured astrocytes, using ouabain-sensitive 86 Rb uptake as an index. l -Glutamate increases glial Na+ ,K+ -ATPase activity in a concentration-dependent manner with an EC50 = 67 µ M . Both l - and d -aspartate, but not d -glutamate, produce a similar response, an observation that is consistent with an uptake-related effect rather than a receptor-mediated one. Under basal conditions, concentration-dependent inhibition of Na+ ,K+ -ATPase activity in astrocytes by ouabain indicates the presence of a single catalytic site with a low affinity for ouabain ( K 0.5 = 113 µ M ), compatible with the presence of an α1 isozyme. On stimulation with glutamate, however, most of the increased activity is inhibited by low concentrations of ouabain ( K 0.5 = 20 n M ), thus revealing a high-affinity site akin to the α2 isozyme. These results suggest that astrocytes possess a glutamate-sensitive isoform of Na+ ,K+ -ATPase that can be mobilized in response to increased neuronal activity. 相似文献
6.
Abstract: Rat brain microsomes were preincubated with S -adenosylmethionine (SAM), MgCl2 , and CaCl2 , then re-isolated, and the activity of Na+ ,K+ -ATPase determined. SAM inhibited the Na+ ,K+ -ATPase activity compared with microsomes subjected to similar treatment in the absence of SAM. A biphasic inhibitory effect was observed with a 50% decrease at a SAM concentration range of 0.4 μ M -3.2 μ M and a 70% reduction at a concentration range above 100 μ M . Inclusion of either S- adenosylhomocysteine or 3-deazaadenosine in the preincubations prevented the SAM inhibition of Na+ ,K+ -ATPase activity. The inhibition by SAM appeared to be Mg2+ - or Ca2+ -dependent. 相似文献
7.
Keiko Mitsunaga-Nakatsubo Miyuki Kanda Ken Yamazaki Hiroto Kawashita Akiko Fujiwara Kazumi Yamada Koji Akasaka Hiraku Shimada Ikuo Yasumasu 《Development, growth & differentiation》1992,34(6):677-684
A marked increase in the Na+ , K+ -ATPase activity of sea urchin embryos occurred following an elevation of its mRNA level, revealed by Northern blotting analysis, in developmental period between the swimming blastula and the late gastrula stage. cDNA clone of Na+ , K+ -ATPase α-subunit, obtained from γgt10 cDNA library of sea urchin gastrulae, was digested with EcoRl ad Hindlll. The obtained 268 bp cDNA fragment, hybridized to a 4.6 Kb RNA, was used as probe for Northern blotting analysis. The level of Na+ , K+ -ATPase mRNA was higher in embryo-wall cell fraction isolated from late gastrulae (ectoderm cells) than the level in the bag fraction, containing mesenchyme cells (mesoderm cells) and archenteron (endoderm cells). The activity of Na+ , K+ -ATPase and the level of its mRNA were higher in animalized embryos obtained by pulse treatment with A23187 for 3 hr, starting at the 8–16 cell stage and were considerably lower in vegetalized embryos induced by 3 hr treatment with Li+ than that in normal embryos at the post gastrula corresponidng stage. Augmentation of Na+ , K+ -ATPase gene expression can be regarded as a marker for ectoderm cell differentiation at the post gastrula stage, which results from determination of cell fate in prehatching period. 相似文献
8.
Susana González Claudia Grillo Alejandro G. De Nicola Gerardo Piroli Jesus Angulo Bruce S. McEwen Alejandro F. De Nicola 《Journal of neurochemistry》1994,63(5):1962-1970
Abstract: The Na+ ,K+ -ATPase plays a key role in the regulation of ion fluxes and membrane repolarization in the CNS. We have studied glucocorticoid effects on biosynthesis of the Na+ ,K+ -ATPase and on ouabain binding in the ventral horn of the spinal cord using intact rats, adrenalectomized (ADX) rats, and ADX rats receiving dexamethasone (ADX + DEX) during 4 days. Cryostat sections from spinal cords were incubated with a 35 S-oligonucleotide coding for the α3 -subunit or a 3 H-cDNA coding for the β1 -subunit of the Na+ ,K+ -ATPase using in situ hybridization techniques. In ventral horn motoneurons, grain density per cell and grain density per area of some for both probes were slightly reduced in ADX rats but significantly increased in the ADX + DEX group, using ANOVA and the Bonferroni's test. Statistical analysis of frequency histograms of neuronal densities further indicated a significant shift to the right for intact rats compared with ADX rats for both probes. Concomitantly, [3 H]ouabain binding to membrane preparations from ventral horns was reduced in ADX rats and restored to normal by DEX administration. No effect of adrenalectomy or DEX treatment was obtained in the dorsal horn. In conclusion, glucocorticoids positively modulate the mRNA for the α3 -subunit and the β1 -subunit of the Na+ ,K+ -ATPase and recover ouabain binding to normal values. The increments of the synthesis and activity of an enzyme affecting membrane repolarization and synaptic neurotransmission are consistent with the alleged stimulatory effect of glucocorticoids on spinal cord function. 相似文献
9.
10.
Abstract: We have previously purified and characterized a nervous system-specific glycoprotein antigen from adult Drosophila heads, designated Nervana [nerve antigen (NRV)] and identified two separate genes coding for three different proteins. All three proteins share homology with the β subunits of Na+ ,K+ -ATPase from various other species. In this study we have isolated a new Drosophila Na+ ,K+ -ATPase α subunit cDNA clone (PSα; GenBank accession no. AF044974) and demonstrate expression of functional Na+ ,K+ -ATPase activity when PSα mRNA is coinjected into Xenopus oocytes along with any of the three different Nrv mRNAs. Western blotting, RNase protection assays, and immunocytochemical staining of adult fly sections indicate that NRV2 is expressed primarily in the nervous system. Staining is most intense in the brain and thoracic ganglia and is most likely associated with neuronal elements. NRV1 is more broadly expressed in muscle and excretory tissue and also shows diffuse distribution in the nervous system. Similar to other species, Drosophila expresses multiple isoforms of Na+ ,K+ -ATPase subunits in a tissue- and cell type-specific pattern. It will now be possible to use the advantages of Drosophila molecular and classical genetics to investigate the phenotypic consequences of altering Na+ ,K+ -ATPase expression in various cell and tissue types. 相似文献
11.
The effects of abscisic acid (ABA) on growth, uptake and translocation of potassium ions, K+ ,Mg2+ -ATPase activity and transpiration were investigated in young wheat ( Triticum aestivum L. cv. Martonvásári-8) plants grown at different K+ supplies. Long-term treatment with ABA (10 μ M ) reduced growth in high-K+ plants, but had less effect under low-K+ conditions. K+ (86 Rb) uptake was inhibited by about 70 and 40% in low- and high-K+ plants, respectively. The stimulation by K+ of the Mg2+ -ATPase activity in the root microsomal fraction was lost with ABA treatment. It is suggested that the inhibitory effect of ABA on K+ uptake may be related to this effects on the K+ ,Mg2+ -ATPase. Translocation of K+ to the shoot was inhibited in low-K+ plants only, and it was not affected in high-K+ plants. In parallel to this, ABA treatment reduced transpiration by about 50% in low-K+ plants, whereas a much smaller effect was seen in high-K+ plants. These observations suggest that the regulation by ABA of the stomatal movements is strongly counteracted by high-K+ status. 相似文献
12.
Goizane Marcaida Elena Kosenko María-Dolores Miñana Santiago Grisolía Vicente Felipo 《Journal of neurochemistry》1996,66(1):99-104
Abstract: In primary cultures of cerebellar neurons glutamate neurotoxicity is mainly mediated by activation of the NMDA receptor, which allows the entry of Ca2+ and Na+ into the neuron. To maintain Na+ homeostasis, the excess Na+ entering through the ion channel should be removed by Na+ ,K+ -ATPase. It is shown that incubation of primary cultured cerebellar neurons with glutamate resulted in activation of the Na+ ,K+ -ATPase. The effect was rapid, peaking between 5 and 15 min (85% activation), and was maintained for at least 2 h. Glutamate-induced activation of Na+ ,K+ -ATPase was dose dependent: It was appreciable (37%) at 0.1 µ M and peaked (85%) at 100 µ M . The increase in Na+ ,K+ -ATPase activity by glutamate was prevented by MK-801, indicating that it is mediated by activation of the NMDA receptor. Activation of the ATPase was reversed by phorbol 12-myristate 13-acetate, an activator of protein kinase C, indicating that activation of Na+ ,K+ -ATPase is due to decreased phosphorylation by protein kinase C. W-7 or cyclosporin, both inhibitors of calcineurin, prevented the activation of Na+ ,K+ -ATPase by glutamate. These results suggest that activation of NMDA receptors leads to activation of calcineurin, which dephosphorylates an amino acid residue of the Na+ ,K+ -ATPase that was previously phosphorylated by protein kinase C. This dephosphorylation leads to activation of Na+ ,K+ -ATPase. 相似文献
13.
Alain Gerbi Marcel Debray Jean-Michel Maixent† Claude Chanez Jean-Marie Bourre 《Journal of neurochemistry》1993,60(1):246-252
Abstract: The Na+ sensitivity of whole brain membrane Na+ ,K+ -ATPase isoenzymes was studied using the differential inhibitory effect of ouabain (α1, low affinity for ouabain; α2, high affinity; and α3, very high affinity). At 100 m M Na+ , we found that the proportion of isoforms with low, high, and very high ouabain affinity was 21, 38, and 41%, respectively. Using two ouabain concentrations (10−5 and 10−7 M ), we were able to discriminate Na+ sensitivity of Na+ , K+ -ATPase isoenzymes using nonlinear regression. The ouabain low-affinity isoform, α1, exhibited high Na+ sensitivity [ K a of 3.88 ± 0.25 m M Na+ and a Hill coefficient ( n ) of 1.98 ± 0.13]; the ouabain high-affinity isoform, α2, had two Na+ sensitivities, a high ( K a of 4.98 ± 0.2 m M Na+ and n of 1.34 ± 0.10) and a low ( K a of 28 ± 0.5 m M Na+ and an n of 1.92 ± 0.18) Na+ sensitivity activated above a thresh old (22 ± 0.3 m M Na+ ); and the ouabain very-high-affinity isoform, α3, was resolved by two processes and appears to have two Na+ sensitivities (apparent K a values of 3.5 and 20 m M Na+ ). We show that Na+ dependence in the absence of ouabain is the result of at least of five Na+ reactivities. This molecular functional characteristic of isoenzymes in membranes could explain the diversity of physiological roles attributed to isoenzymes. 相似文献
14.
Sune Pettersson 《Physiologia plantarum》1986,66(1):122-128
Plants of barley ( Hordeum vulgare L. cv. Salve) were grown with 6.5–35% relative increase of K+ supply per day (RKR) using a special computer-controlled culture unit. After a few days on the culture solution the plants adapted their relative growth rate (RGR) to the rate of nutrient supply. The roots of the plants remained in a low salt status irrespective of the rate of nutrient supply, whereas the concentration of K+ in shoots increased with RKR. Both Vmax and Km for K+ (86 Rb) influx increased with RKR. It is concluded that with a continuous and stable K+ stress, the K+ uptake system is adjusted to provide an effective K+ uptake at each given RKR. Allosteric regulation of K+ influx does not occur and efflux of K+ is very small. 相似文献
15.
Abstract. Rates of proton extrusion and potassium (86 Rb) influx by intact roots of barley ( Hordeum vulgare cvs . Fergus, Conquest and Betzes) plants were simultaneously measured in short-term (15min) experiments. The nature and extent of apparent coupling between these ion fluxes was explored by manipulating conditions of temperature, pH and cation composition and concentration during flux determinations. In addition, the influence of salt status upon these fluxes was examined. At low K+ concentrations (0.01 to 1 mol m−3 ), H+ efflux and K+ influx were strongly correlated in both low- and high-K+ roots, although K+ : H+ exchange stoichiometries were almost consistently greater than 2:1. At higher concentrations (1 to 5 mol m−3 ), H+ efflux was either reduced or remained unchanged while K+ influxes increased. In the presence of Na2 SO4 , rates of H+ extrusion demonstrated similar cation dependence, although below 10 mol m−3 Na2 SO4 , H+ fluxes were generally 50% lower than in equivalent concentrations of K2 SO4 . These observations are considered in the context of current hypotheses regarding the mechanisms of k+ /H+ exchange. 相似文献
16.
Renato S. Gomez Marcus Vinicius Gomez Marco Antônio M. Prado 《Journal of neurochemistry》1996,66(4):1440-1447
Abstract: Ouabain, an Na+ ,K+ -ATPase inhibitor, increases the release of acetylcholine (ACh) from various preparations in a Ca2+ -independent way. However, in other preparations the release of ACh evoked by ouabain is dependent on the presence of extracellular calcium. In the present study, we have labeled the ACh of myenteric plexus longitudinal muscles of guinea pig ileum and compared the effect of calcium channel blockers on ouabain-evoked release of [3 H]ACh. Release of [3 H]ACh evoked by ouabain is dose dependent and decreased markedly in the absence of calcium or in the presence of cadmium, a nonspecific calcium channel blocker. N-type calcium channel blockage by the ω-conotoxins GVIA (selective N-type calcium channel blocker) and MVIIC (a nonselective calcium channel blocker) inhibited by 45 and 55%, respectively, the release of [3 H]ACh. L-type calcium channel suppression by low concentrations of verapamil, nifedipine, and diltiazem had no effect on the release of [3 H]ACh. The release of transmitter was also not affected significantly by nickel, a T-type calcium channel blocker. In addition, ω-agatoxin-IVA, at concentrations that block P- and Q-type calcium channels, did not affect significantly the release of [3 H]ACh. Thus, extracellular Ca2+ is essential for the release of ACh induced by ouabain from guinea pig ileum myenteric plexus. In this preparation, the N-type calcium channel plays a dominant role in transmitter release evoked by inhibition of Na+ ,K+ -ATPase, but other routes of calcium entry in addition to these channels can also support the release of neurotransmitter induced by ouabain. 相似文献
17.
Abstract Washed cells of Rhodopseudomonas sphaeroides forma sp. denitrificans , grown under photodenitrifying conditions, exhibited K+ uptake dependent on the transmembrane proton gradient (Δ pH). These cells also acidified the suspension medium in response to K+ pulses both aerobically and anaerobically in light and in the dark. The results indicate that the photodenitrifier has a reversible K+ /H+ exchange activity which reflects its role in regulating the intracellular K+ concentration, as well as intracellular pH. The acidification of the external medium resulting from K+ pulses was inhibited by carbonyl cyanide- m -chlorophenylhydrazone (CCCP) indicating that the antiporter is energy-dependent. Addition of KCl to washed cells depolarized the membrane potential (Δψ) with a concomitant increase in ΔpH, indicating that the K+ /H+ antiporter was electrogenic. 相似文献
18.
Sune Pettersson 《Physiologia plantarum》1981,52(4):431-436
Passive fluxes of K+ (86 Rb) into roots of sunflower ( Helianthus annuus L. cv. Uniflorus) were determined at low K+ concentration (0.1 and 1.0 mM K+ ) in the ambient solution. Metabolic uptake of K+ was inhibited by 10−4 M 2,4-dinitrophenol (DNP). K+ (86 Rb) fluxes were studied both continuously and by time differentiation of uptake. In high K+ roots passive uptake was directly proportional to the K+ concentration of the uptake solution, indicating free diffusion. This assumption was supported by the fact that passive Rb+ uptake was not affected by high K+ concentrations. In low K+ roots the passive uptake of K+ was higher than in high K+ roots. The increase was possibly due to carrier-mediated K+ transport. As K+ effluxes were quantitatively similar to influxes, it is suggested that passive K+ fluxes represent exchange diffusion without relation to net K+ transport. 相似文献
19.
Rupinder Kaur Kanwar N.K. Ganguly J.R. Kanwar Lata Kumar B.N.S. Walia 《FEMS microbiology letters》1994,124(3):381-385
Abstract Unidirectional fluxes of Na+ , Cl− and 3-O-methyl-D-glucose (3-MG) were measured in vitro across Campylobacter jejuni live culture-infected and control rat ileal short-circuited tissues by the Using Chamber technique. Net secretion of Na+ and enhanced secretion of Cl− ions was observed in the infected animals ( P < 0.001, n =6) as compared to the net absorption of Na+ and marginal secretion of Cl− ions in the control animals. There was a significant decrease in the mucosal-to-serosal fluxes of 3-MG in C. jejuni -infected rat ileum. The specific Na+ ,K+ -ATPase activity when measured biochemically in the membrane-rich fraction of enterocytes was found to be significantly lower (58%) in the infected group as compared to the control group ( P < 0.001). Our results therefore suggest that infection with an enterotoxigenic C. jejuni inhibits the Na+ ,K+ -ATPase activity in rat enterocytes. The impairment of Na+ ,K+ -ATPase activity thus appears to induce a secondary change in Na+ ,Cl− and 3-MG transport in vitro in rat ileum. 相似文献
20.
The metabolic response of juvenile coho salmon Oncorhynchus kisutch to different salinities was examined, using whole-animal oxygen consumption rates and gill Na+ , K+ -ATPase activities as indicators of osmoregulatory energetics. Coho salmon smolts were acclimated to fresh water (FW), isosmotic salinity (ISO, 10‰) and sea water (SW, 28‰) and were sampled for up to 6 weeks for plasma levels of cortisol, glucose and ions (Na+ , K+ , Cl− ), gill Na+ , K+ -ATPase activity and oxygen consumption rates. Following an initial adjustment period, plasma constituents in SW fish returned to near-FW values, indicating that the fish were acclimated to SW by day 21. Gill Na+ , K+ -ATPase activities on days 21 and 42 were lowest in ISO, higher in FW and highest in SW. This result is consistent with the idea that less energy would be required to maintain ion balance in an isosmotic environment, where the ionic gradients between extracellular fluid and water would be minimal. Oxygen consumption rates of swimming fish (1 body length s−1 ), however, did not differ significantly between the three test salinities after 6 weeks. The results of this study suggest that the metabolic response of juvenile salmonids to changes in salinity is dependent on life-history stage (e.g. fry v . smolt), and that oxygen consumption rates do not necessarily reflect osmoregulatory costs. 相似文献