共查询到20条相似文献,搜索用时 15 毫秒
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In order to study the role of glutamine from glial cells for the synthesis of transmitter amino acids, the effect of the gliotoxic substance fluorocitrate on amino acid release from slices was investigated. In vivo treatment with 1 nmol fluorocitrate reduced the Ca2+ dependent K+ evoked release of endogenous glutamate and GABA from the slices, whereas the glutamine efflux decreased and alanine efflux increased. The K+ evoked release of [3H]d-aspartate increased during fluorocitrate treatment. The latter is consistent with an inhibited uptake ofd-aspartate into glial cells. Incubation of striatal slices with fluorocitrate (0.1 mM) decreased the glutamine efflux and increased the alanine efflux. Similar to the in vivo condition, fluorocitrate increased the K+ evoked [3H]d-asparate release, but the K+ evoked release of endogenous glutamate and GABA increased rather than decreased. The ratio between the K+ evoked release of exogenousd-aspartate to endogenous glutamate increased in both cases. The results suggest an important role of glial cells in the synthesis and inactivation of transmitter amino acids.Special Issue dedicated to Prof. Holger Hydén. 相似文献
3.
The transport of [3H]2-deoxyglucose by brain slices was studied. Cerebral cortex slices were incubated in vitro in the presence of [3H]2-deoxyglucose, orl-[3H] glucose as a marker for diffusion. Transport was defined as the difference between [3H]2DG uptake andl-[3H]glucose uptake. Half-maximal velocity was seen at 2.0 mM 2DG and [3H]2DG transport was not inhibited by 20-fold higher concentrations ofl-glucose. Net [3H]2DG transport was unchanged in media deficient in Na+, K+, Mg2+, Ca2+ or Cl–. Uptake was significantly inhibited by 1.0 mM 2,4-DNP and a suggestion of inhibition by azide was seen. These data are consistent with a hypothesis that hexose transport in the brain depends to some extent upon mitochondrial energy. 相似文献
4.
The human erythroleukaemic cell line K562, in response to various chemical agents, undergoes differentiation and exhibits exclusive production of fetal and embryonic haemoglobins. In this study we have compared the efficiency of natural growth factors interleukin-3 and erythropoietin and three chemical inducers such as dimethyl sulfoxide (DMSO, 1.9%), phorbol-12-myristate-13-acetate (PMA, 50 ng/ml) and hemin (25 μm) on growth and differentiation of these cells. Erythropoietin significantly stimulated the growth of K562 cells (P<0.0001), while interleukin-3 did not (P= 0.2783). However, neither of these growth factors individually or together induced differentiation of K562 cells. Hemin appears to be more efficient than DMSO or PMA in differentiation of K562 cells as measured by benzidine positive cells (70% or more). The differentiation of K562 cells by hemin occurs independently of protein kinase-C activation and the arrest of DNA synthesis. In contrast, hemin significantly stimulated RNA and protein synthesis (P<0.0001) as measured by [3H]-uridine and [3H]-leucine incorporation respectively. Analysis of hemin-treated K562 nuclear extract on sodium dodecylsulphate gel electrophoresis showed that one protein band of molecular weight 70 kDa decreased after 48 h of incubation in the presence of 25 μm hemin. The disappearance of this protein can be prevented by cycloheximide (100 μg/ml) and actinomycin D (0.1 μg/ml) and thus indicating that the removal of 70 kDa protein seems to be dependent on RNA and protein synthesis. The regulatory role of 70 kDa protein in hemin-induced differentiation of K562 cells is discussed. 相似文献
5.
Effect of Nipecotic Acid, a γ-Aminobutyric Acid Transport Inhibitor, on the Turnover and Release of γ-Aminobutyric Acid in Rat Cortical Slices 总被引:11,自引:11,他引:0
John C. Szerb 《Journal of neurochemistry》1982,39(3):850-858
Abstract: To see the effect of a γ-aminobutyric acid GABA uptake inhibitor on the efflux and content of endogenous and labeled GABA, rat cortical slices were first labeled with [3H]GABA and then superfused in the absence or presence of 1 mM nipecotic acid. Endogenous GABA released or remaining in the slices was measured with high performance liquid chromatography, which was also used to separate [3H]GABA from its metabolites. In the presence of 3 mM K+, nipecotic acid released both endogenous and [3H]GABA, with a specific activity four to five times as high as that present in the slices. The release of labeled metabolite(s) of [3H]GABA was also increased by nipecotic acid. The release of endogenous GABA evoked by 50 mM K+ was enhanced fourfold by nipecotic acid but that of [3H]GABA was only doubled when expressed as fractional release. In a medium containing no Ca2+ and 10 mM Mg2+, the release evoked by 50 mMK+ was nearly suppressed in either the absence or the presence of nipecotic acid. In the absence of nipecotic acid electrical stimulation (bursts of 64 Hz) was ineffective in evoking release of either endogenous or [3H]GABA, but in the presence of nipecotic acid it increased the efflux of endogenous GABA threefold, while having much less effect on that of [3H]GABA. Tetrodotoxin (TTX) abolished the effect of electrical stimulation. Both high K+ and electrical stimulation increased the amount of endogenous GABA remaining in the slices, and this increase was reduced by omission of Ca2+ or by TTX. The results suggest that uptake of GABA released through depolarization is of major importance in removing GABA from extracellular spaces, but the enhancement of spontaneous release by nipecotic acid may involve intracellular heteroexchange. Depolarization in the presence of Ca2+ leads to an increased synthesis of GABA, in excess of its release, but the role of this excess GABA remains to be established. 相似文献
6.
Tetsuhiro Sakai Toshitaka Ikehara Hiroshi Miyamoto Kenichi Kaniike 《Life sciences》1981,29(23):2429-2436
Growth of HeLa cells cultured with a chemically defined medium was slightly stimulated in the presence of 5% dialyzed calf serum. The growth-promoting action of serum was more conspicuous when cell growth was suppressed in the same medium, in which K+ was replaced by Rb+ to various ratios. The growth-promoting factors(s) of serum was heat-labile. Upon addition of dialyzed serum, passive K+ or Rb+ influx was increased, whereas the active cation uptake was unaffected and cell K+ was rather decreased. The serum did not alter uptake of [3H] amino acids. Also, protein synthesis inhibited in the Rb+-substituted medium was not stimulated significantly, except that observed only when the external K+/Rb+ ratio was . From the distinct effects of serum on cell growth and protein synthesis, we conclude that (i) the serum-induced stimulation of cell growth, which is suppressed in the Rb+-substituted medium, is not a result of the direct effect of serum on synthesis of bulk protein, but a reflection of the effect on another mechanism(s) required for cell growth; and that (ii) this action is basically identical with the growth-promoting action on cells cultured in the normal medium. 相似文献
7.
M. Szamel J. Somogyi I. Csuks F. Solymosy 《Biochimica et Biophysica Acta (BBA)/General Subjects》1980,633(3):347-360
Ouabain inhibited in a concentration-dependent and completely reversible way, the synthesis of DNA, RNA and protein in phytohemagglutinin and concanavalin A-stimulated human lymphocytes without affecting the uptake of nucleosides and amino acids into the cells. On the other hand, ouabain even at very high concentrations was unable to interfere with the binding of [3H]concanavalin A. No correlation was found between the inhibition by ouabain of macromolecular synthesis and that of K+ transport. The inhibitor effect of ouabain on the stimulation of macromolecular synthesis could be partially reversed by higher concentrations of K+, due to the direct inhibition of ouabain binding. Ouabain added to the cultures at different stages of cell growth suppressed the incorporation of thymidine to various extents. Both ouabain sensitive stages fell in a period preceding the onset of mitosis and were characterized by very active thymidine incorporation. Lymphocytes were most sensitive to ouabain within the S phase. The results suggest that ouabain interferes with mitogen-triggered membrane-associated events, other than K+ transport, controlling mitosis at distinct phases of the cell cycle. 相似文献
8.
Franca Cerrito Gianna Casazza Giulio Levi Maurizio Raiteri 《Neurochemical research》1980,5(2):115-121
The aim of the present study was to compare the release pattern of [3H]dopamine ([3H]DA) originated from [3H]tyrosine or by uptake in striatal synaptosomes. Synaptosomes prelabeled either with [3H]DA or with [3H]tyrosine were superfused in three conditions stimulating DA release by different mechanisms: (1) depolarization with high K+; (2) inversion of the Na+ gradient across the plasma membrane; (3) exposure tod-amphetamine. Since DA contained in different pools may exit from nerve endings by different processes, DA release was analyzed in the presence or in the absence of nomifensine which allows discrimination between carrier-mediated and carrier-independent processes. The pattern of DA release in the three conditions tested was identical, whether [3H]DA originated from synthesis or from uptake. Nomifensine did not affect the high-K+-induced release and inhibited that induced by the other two stimuli. The results suggest that newly synthesized and recaptured DA have a similar compartmentation in nerve endings. 相似文献
9.
Ionic and Metabolic Requirements for High-Affinity Choline Uptake and Acetylcholine Synthesis in Nerve Terminals at a Neuromuscular Junction 总被引:2,自引:1,他引:1
Abstract: We have shown previously that in the chick ciliary nerve-iris muscle preparation Na+-dependent high-affinity choline uptake was confined to the nerve terminals. In this paper the sodium-dependent high-affinity choline uptake (SDHACU), which is coupled to acetylcholine (ACh) synthesis, was further characterized by measuring uptake of [3H]choline and its conversion to [3hjach under a variety of ionic and metabolic perturbations. Mannitol equilibration with the extracellular space was found to occur in less than 1 min in this preparation. Na+-dependent choline (Ch+) uptake was shown to be linear for 16 min and to reach an equilibrium before Na+-independent Ch+ uptake, which continued to increase for 60 min. Elevated [K+]0 concentrations inhibited Ch+ uptake and ACh synthesis. Glycolytic and respiratory inhibitors also reduced both processes, as did ouabain and omission of [K+]0. Incubation conditions that reduce transmitter release had no effect on inhibition by high [K+]0. Reduction of SDHACU and sodium-dependent ACh synthesis by depolarization with high [K+]0 or by inhibition of Na, K-ATPase implies that the electrochemical gradients for Ch+ and Na+ are important in providing a driving force for high-affinity Ch+ uptake. The inhibition by metabolic blockers suggests active transport, but the effects may be indirect, caused by reduced Na, K-ATPase activity and alterations in membrane potential. While most metabolic inhibitors exerted parallel effects on both Ch+ uptake and ACh synthesis, in some cases Ch+ uptake was more strongly inhibited than ACh synthesis. This occurred in preparations incubated with high [K+]0 and ouabain. Na+-dependent Ch+ uptake and ACh synthesis were found to be temperature-dependent with a Q10 (20–30°) of 3.6 and 6.6, respectively and a Q10 (30–40°) of 1.3 and 1.0, respectively. Inhibition of acetylcholinesterase by paraoxon increases to 92% the proportion of the Ch+ taken up which is converted to ACh. ACh did not reduce Ch+ transport when present at 100 μM. 相似文献
10.
The uptake of [3H]actinomycin D ([3H]AD) by ConA-stimulated lymphocytes was followed during 96 h of incubation and correlated with the level of nuclear proteins in the nucleus, DNA synthesis and the degree of AD-induced inhibition of RNA and DNA synthesis. During the first 48 h there is a parallel increase of drug binding to cells and a rising level of non-histone proteins (NHP) in the nucleus. During the next 48 h, DNA synthesis occurs, drug uptake decreases and the nuclear level of NHP continues to rise. The level of histones remains constant during 96 h. The variations in cellular [3H]AD uptake during 96 h are not due to changes in cell membrane permeability, since similar variations in drug binding are observed in isolated cell nuclei. NHP, obtained as 0.25 M NaCl extracts of cell nuclei, increase binding of [3H]AD to nuclei isolated from non-stimulated lymphocytes, while histones have no such effect. NHP extracted with phenol, after washing the nuclei with salt and acid solutions, or extracted with 0.25 M NaCl from non-stimulated and stimulated lymphocytes and Chang liver cells are equally active to bind [3H]AD to nuclei of non-stimulated lymphocytes. NHP from Chang cells, purified by DNA-cellulose chromatography using calf thymus DNA, stimulated [3H]AD binding to lymphocyte nuclei, indicating that the drug-binding activity is due to proteins binding to DNA. NHP increase binding of [3H]AD to pure DNA in the absence of histones. The degree of [3H]AD binding to ConA-stimulated lymphocytes during 96 h correlated with the degree of inhibition of RNA and DNA synthesis by AD. 相似文献
11.
The uptake of [3H]actinomycin D ([3H]AD) by ConA-stimulated lymphocytes was followed during 96 h of incubation and correlated with the level of nuclear proteins in the nucleus, DNA synthesis and the degree of AD-induced inhibition of RNA and DNA synthesis. During the first 48 h there is a parallel increase of drug binding to cells and a rising level of non-histone proteins (NHP) in the nucleus. During the next 48 h, DNA synthesis occurs, drug uptake decreases and the nuclear level of NHP continues to rise. The level of histones remains constant during 96 h. The variations in cellular [3H]AD uptake during 96 h are not due to changes in cell membrane permeability, since similar variations in drug binding are observed in isolated cell nuclei. NHP, obtained as 0.25 M NaCl extracts of cell nuclei, increase binding of [3H]AD to nuclei isolated from non-stimulated lymphocytes, while histones have no such effect. NHP extracted with phenol, after washing the nuclei with salt and acid solutions, or extracted with 0.25 M NaCl from non-stimulated and stimulated lymphocytes and Chang liver cells are equally active to bind [3H]AD to nuclei of non-stimulated lymphocytes. NHP from Chang cells, purified by DNA-cellulose chromatography using calf thymus DNA, stimulated [3H]AD binding to lymphocyte nuclei, indicating that the drug-binding activity is due to proteins binding to DNA. NHP increase binding of [3H]AD to pure DNA in the absence of histones. The degree of [3H]AD binding to ConA-stimulated lymphocytes during 96 h correlated with the degree of inhibition of RNA and DNA synthesis by AD. 相似文献
12.
Joan M. V. Blasi Valentín Ceña Carmen González-García Jordi Marsal Carles Solsona 《Neurochemical research》1988,13(11):1035-1041
We have studied the correlation between [3H]ouabain binding sites, (Na++K+)ATPase (EC 3.6.1.3) activity and acetylcholine (ACh) release in different subcellular fractions ofTorpedo marmorata electric organ (homogenate, synaptosomes, presynaptic plasma membranes). Presynaptic plasma membranes contained the greater number of [3H]ouabain binding sites in good agreement with the high (Na++K+)ATPase activity found in this fraction. Blockade of this enzymatic activity by ouabain dose-dependently induced ACh release from pure cholinergic synaptosomes, either in the presence or absence of extracellular calcium ions. We suggest that one of the mechanisms involved in the ouabain-induced ACh release in the absence of Ca2+
o may be an increase in Na+
i that could (a) evoke Ca2+ release from internal stores and (b) inhibit ATP-dependent Ca2+ uptake by synaptic vesicles. 相似文献
13.
J. B. Lombardini 《Neurochemical research》1993,18(2):193-202
The effects of spontaneous and evoked [3H]taurine release from a P2 fraction prepared from rat retinas were studied. The P2 fraction was preloaded with [3H]taurine under conditions of high-affinity uptake and then examined for [3H]taurine efflux utilizing superfusion techniques. Exposure of the P2 fraction to high K+ (56 mM) evoked a Ca2+-independent release of [3H]taurine. Li+ (56 mM) and veratridine (100 M) had significantly less effect (8–15% and 15–30%, respectively) on releasing [3H]taurine compared to the K+-evoked release. 4-Aminopyridine (1 mM) had no effect on the release of [3H]taurine. The spontaneous release of [3H]taurine was also Ca2+-independent. When Na+ was omitted from the incubation medium K+-evoked [3H]taurine release was inhibited by approximately 40% at the first 5 minute depolarization period but was not affected at a second subsequent 5 minute depolarization period. The spontaneous release of [3H]taurine was inhibited by 60% in the absence of Na+. Substitution of Br– for Cl– had no effect on the release of either spontaneous or K+-evoked [3H]taurine release. However, substitution of the Cl– with acetate, isethionate, or gluconate decreased K+-evoked [3H]taurine release. Addition of taurine to the superfusion medium (homoexchange) resulted in no significant increase in [3H]taurine efflux. The taurine-transport inhibitor guanidinoethanesulfonic acid increased the spontaneous release of [3H]taurine by approximately 40%. These results suggest that the taurine release of [3H]taurine is not simply a reversal of the carrier-mediated uptake system. It also appears that taurine is not released from vesicles within the synaptosomes but does not rule out the possibility that taurine is a neurotransmitter. The data involving chloride substitution with permeant and impermeant anions support the concept that the major portion of [3H]taurine release is due to an osmoregulatory action of taurine while depolarization accounts for only a small portion of [3H]taurine release. 相似文献
14.
L. C. Rai M. Raizada N. Mallick Yashmin Husaini A. K. Singh S. K. Dubey 《Biometals》1990,2(4):229-234
Summary This study presents the effects of Cr, Pb, Ni and Ag on growth, pigments, protein, DNA, RNA, heterocyst frequency, uptake of NH4
+ and N03
–, loss of electrolytes (Na+ and K+), nitrate reductase and glutamine synthetase activities ofNostoc muscorum. The statistical tests revealed a direct positive correlation between the metal concentration and inhibition of different processes. Ni was found to be more toxic against growth, pigments and heterocyst differentiation compared to the other metals. Inhibition of pigment showed the following trend: chlorophyll > phycocyanin > carotenoid. No generalized trend for inhibition of macromolecules was observed. The loss of K+ and Na+ as affected by Cr, Ni and Pb was similar but more pronounced for K+ than Na+. The inhibition of physiological variables depicted the following trend: Na+ loss > K+ loss > glutamine synthetase > NH4 uptake > growth > N03
– uptake > nitrate reductase > heterocyst frequency. This study therefore suggests that loss of electrolytes can be used as a first signal of metal toxicity in cyanobacteria. However, further study is needed to confirm whether the abnormality induced by nickel (branch formation) is a physiological or genetic phenomenon. 相似文献
15.
《Neurochemistry international》1987,11(2):183-188
Acetylcholine enhanced in a concentration-dependent way the K+ (15 mM)-evoked release of [3H]dopamine from synaptosomes isolated from rat corpus striatum and prelabeled with the radioactive catecholamine. The concentration-effect curve of ACh obtained in presence of 1.2 mM Ca2+ was progressively shifted to the left when [Ca2+] was lowered to 0.4 and to 0.2 mM. Intrastriatal injections of kainic acid reduced (70%) the uptake of [3H]choline in synaptosomes prepared 8 days after the lesion but did not affect significantly the uptake of [3H]dopamine. Also the release of [3H]dopamine evoked by K+ was minimally affected by kainic acid treatment. In contrast, acetylcholine (tested in presence of 1.2 or 0.2 mM Ca2+) was much more effective in enhancing [3H]dopamine release in synaptosomes from kainic acid-lesioned than from unlesioned striata. The results suggest that muscarinic receptors located on dopamine nerve terminals undergo supersensitivity following intrastriatal kainic acid injection. 相似文献
16.
Donna L. Wolcott 《Experimental cell research》1982,137(2)
Vegetalization of sea urchin embryos by Li+ is characterized by rates of protein synthesis which are normal during cleavage, and decline after hatching. This paper tests the hypothesis that Li+ interferes with RNA synthesis during cleavage, resulting in the decline in protein synthesis at hatching when newly synthesized mRNA becomes critical for further normal development. Treatment with Li+ does cause a decline in the incorporation of [3H]guanosine into RNA. However, this decline could be accounted for by reduced uptake of the labeled precursor with a concomitant reduction in precursor pool specific activity. Therefore, reduced protein synthesis after hatching in Li+-treated embryos cannot be accounted for by a comparable reduction in RNA synthesis. 相似文献
17.
The effect of serotonin agonists on the depolarization (K+)-induced, calcium-dependent, release of [3H]dopamine (DA) from rat nucleus accumbens and striatal slices was investigated. Serotonin enhanced basal3H overflow and reduced K+-induced release of [3H]DA from nucleus accumbens slices. The effect of serotonin on basal3H overflow was not altered by the serotonin antagonist, methysergide, or the serotonin re-uptake blocker, chlorimipramine, but was reversed by the DA re-uptake carrier inhibitors nomifensine and benztropine. With the effect on basal overflow blocked, serotonin did not modulate K+-induced release of [3H]DA in the nucleus accumbens or striatum. The serotonin agonists, quipazine (in the presence of nomifensine) and 5-methoxytryptamine, did not significantly affect K+-induced release of [3H]DA in the nucleus accumbens. This study does not support suggestions that serotonin receptors inhibit the depolarization-induced release of dopamine in the nucleus accumbens or striatum of the rat brain. The present results do not preclude the possibility that serotonin may affect the mesolimbic reward system at a site which is post-synaptic to dopaminergic terminals in the nucleus accumbens. 相似文献
18.
Francesca Oppedisano 《生物化学与生物物理学报:生物膜》2008,1778(10):2258-2265
Na+ dependent [3H]glutamine uptake was found in liposomes reconstituted with solubilized rat kidney brush border in the presence of intraliposomal K+. The reconstituted system was optimised with respect to the critical parameters of the cyclic detergent removal procedure, i.e., the detergent used for the solubilization, the protein concentration, the detergent/phospholipid ratio and the number of passages through a single Amberlite column. Time dependent [3H]glutamine accumulation in proteoliposomes occurred only in the presence of external Na+and internal K+. The transporter showed low if there is any tolerance towards the substitution of Na+ or K+ for other cations. Valinomycin strongly stimulated the transport indicating that it is electrogenic. Intraliposomal glutamine had no effect. From the dependence of the transport rate on the Na+ concentration cooperativity index close to 1 was derived, indicating that 1 Na+ should be involved in the cotransport with glutamine. The electrogenicity of the transport originated from the Na+ transport. Optimal rate of 0.1 mM [3H]glutamine uptake was found in the presence of 50 mM intraliposomal K-gluconate. At higher K-gluconate concentrations the transport rate decreased. The activity of the reconstituted transporter was pH dependent with optimal function in the range pH 6.5-7.0. [3H]glutamine (and [3H]leucine) uptake was inhibited by all the neutral but not by the positively or negatively charged amino acids. The sulfhydryl reagents HgCl2, mersalyl, p-hydroxymercuribenzoate and the substrate analogue 2-aminobicyclo[2,2,1]heptane-2-carboxylate strongly inhibited the transporter, whereas the amino acid analogue α-(methylamino)isobutyrate had no effect. The inhibition by mersalyl was protected by the presence of the substrate. On the basis of the Na+ dependence, the electrogenic transport mode and the specificity towards the amino acids, the reconstituted transporter was classified as B°-like. 相似文献
19.
《Neurochemistry international》1986,8(2):171-177
Rat hippocampal synaptosomes were used to investigate the effects of muscarinic cholinergic drugs on the release of [3H]noradrenaline and the rate of noradrenaline synthesis. Oxotremorine and acetylcholine caused depression of K+-evoked release of [3H]noradrenaline; these effects were reversed by atropine and pirenzepine. Muscarinic agonists depressed 2-chloroadenosine- and isoprenaline-stimulated noradrenaline synthesis but had no effect on 8-Br-cyclic AMP-stimulated synthesis. Oxotremorine also depressed the K+-acceleration of noradrenaline synthesis. The action of pirenzepine suggests that the inhibition of release and synthesis are mediated by separate muscarinic receptor subtypes. 相似文献
20.
Abstract: Primary astrocyte cultures from neonatal rat brains show uptake of [3H]norepinephrine ([3H]NE). This uptake has a high-affinity component with an apparent Km of approximately 3 × 10?7 M. At 10?7 M [3H]NE both the initial rate of uptake and steady-state content of [3H]NE is inhibited by up to 95% by omission of external Na+. The Na+-dependent component of this uptake is totally inhibited by the tricyclic antidepressants desipramine (DMI) and amitryptyline with IC50 values of 2 × 10?9 and 4 × 10?8 M, respectively. Inhibition of [3H]NE uptake by DMI shows competitive kinetics. These characteristics are essentially identical to those found for high-affinity uptake of NE in total membrane or synaptosome fractions from rodent brains and suggests that such uptake in neural tissue is not exclusively neuronal. 相似文献