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1.
The effects of reserpine and imipramine on intact human platelets have been investigated, utilizing brief thrombin treatment to evaluate serotonin (5HT) uptake into and loss from the vesicular (thrombin-releasable) compartment. Less than five seconds after its addition, reserpine (10?6M) almost completely inhibited the uptake of 5HT into storage vesicles; but induced an outward flux of stored 5HT from vesicles only after more than two minutes following its addition. Imipramine (10?6M), acting over a 30-minute period, caused no loss of vesicular 5HT, but acted within five minutes to inhibit markedly the movement of cytoplasmic 5HT into storage vesicles. It thus seems likely that in human platelets, inhibition of vesicular 5HT uptake does not necessarily lead to the loss of vesicular 5HT. 相似文献
2.
Increased acoplasmic transport of H3-dopamine in nigro-neostriatal neurons after reserpine 总被引:1,自引:0,他引:1
Recent evidence suggests that dopamine can undergo axoplasmic transport in nigro-neostriatal neurons by binding to amine storage granules. In the present experiments it was demonstrated that reserpine pretreatment (10 mg/kg) 24 hours before stereotaxic injections of 3H-DOPA or 3H-dopamine into the substantia nigra increases the amount of 3H-dopamine transported to the neostriatum by about 300 percent. The activity recovered from the substantia nigra was significantly reduced by reserpine pretreatment however. Stereotaxic injection of 14C-leucine into the substantia nigra indicated that neither fast nor slow axoplasmic transport of protein was influenced by reserpine pretreatment in these same neurons. The increased transport of dopamine appears therefore to be due to a relatively selective action of reserpine. The results suggest that reserpine either (i) increases the binding of dopamine to newly synthesized amine storage granules, (ii) increases the number of newly synthesized amine storage granules, or (iii) accelerates the rate of transport of amine storage granules. In addition, the results support the view that reserpine can increase the membrane permeability of adrenergic neurons to the outward movement of catecholamines. 相似文献
3.
During K+ depletion of a mutant of which cannot concentrate this cation, protein synthesis is inhibited but RNA formation continues. The RNA produced during K+ depletion was analyzed by gel electrophoresis. It was found that 4S, 5S and 23S RNA were synthesized by K+-depleted cells whether uninfected or infected with phage T4. In addition, an RNA species moving close to 16S (presumably 17S) and material of about 6–10S were made during K+ depletion. These species of RNA were not evident in growing cells. Methylation of RNA is severely inhibited during K+ depletion. 相似文献
4.
《生物化学与生物物理学报:生物膜》2023,1865(7):184197
Neurotransmitter release from sympathetic terminals is a key avenue for heart regulation. Herein, presynaptic exocytotic activity was monitored in mice atrial tissue using a false fluorescent neurotransmitter FFN511, a substrate for monoamine transporters. FFN511 labeling had similarity with tyrosine hydroxylase immunostaining. High [K+]o depolarization caused FFN511 release, which was augmented by reserpine, an inhibitor of neurotransmitter uptake. However, reserpine lost the ability to increase depolarization-induced FFN511 unloading after depletion of ready releasable pool with hyperosmotic sucrose. Cholesterol oxidase and sphingomyelinase modified atrial membranes, changing in opposite manner fluorescence of lipid ordering-sensitive probe. Plasmalemmal cholesterol oxidation increased FFN511 release upon K+-depolarization and more markedly potentiated FFN511 unloading in the presence of reserpine. Hydrolysis of plasmalemmal sphingomyelin profoundly enhanced the rate of FFN511 loss due to K+-depolarization, but completely prevented potentiating action of reserpine on FFN511 unloading. If cholesterol oxidase or sphingomyelinase got access to membranes of recycling synaptic vesicles, then the enzyme effects were suppressed. Hence, a fast neurotransmitter reuptake dependent on exocytosis of vesicles from ready releasable pool occurs during presynaptic activity. This reuptake can be enhanced or inhibited by plasmalemmal cholesterol oxidation or sphingomyelin hydrolysis, respectively. These modifications of plasmalemmal (but not vesicular) lipids increase the evoked neurotransmitter release. 相似文献
5.
A soluble rat liver nuclear extract containing total RNA polymerase activities also exhibits appreciable amounts of protein kinase activity. This unfractionated protein kinase catalyzes the phosphorylation of both endogenous proteins and exogenous lysine-rich histone in the presence of [γ-32P]ATP and Mg2+. The optimal concentration of Mg2+ is 5 mm for histone phosphorylation and 25 mm for the phosphorylation of endogenous proteins. Cyclic AMP has no effect on the phosphorylation of lysine-rich histone by this unfractionated nuclear protein kinase. However, addition of cyclic AMP causes a reduction in the 32P-labeling of an endogenous protein (CAI) which can be characterized by its mobility during SDS-acrylamide gel electrophoresis and elution in the unbound fraction of a DEAESephadex column. If CAI is first labeled with 32P and then incubated with 10?6m cyclic AMP under conditions where protein kinase activity is inhibited, the presence of the cyclic nucleotide causes a loss of the 32P-labeling of this protein, implying the activation of a substrate-specific protein phosphatase. When rat liver RNA polymerases are purified by DEAE-Sephadex chromatography, protein kinase activity is found in the unbound fraction and in those column fractions containing RNA polymerase I and II. The fractionated protein kinases exhibit different responses to cyclic AMP, the unbound protein kinase being stimulated and the RNA polymerase-associated protein kinases being dramatically inhibited. A second protein (CAII) whose phosphorylated state is modified by cyclic AMP is found within the DEAE-Sephadex column fractions containing RNA polymerase II. The cyclic nucleotide in this case appears to reduce labeling of CAII by inhibition of the protein kinase activity which co-chromatographs with both CAII and RNA polymerase II. Based on molecular weight estimates, neither CAI nor CAII appears to be an RNA polymerase subunit. The identity of CAI as a protein factor whose phosphorylated state influences nuclear RNA synthesis is suggested by the fact that addition of fractions containing CAI to purified RNA polymerase II inhibits the activity of this enzyme, but only if CAI has been previously incubated in the presence of cyclic AMP. 相似文献
6.
The rate of synthesis of RNA was studied under salt and water stresses created by the addition of four different salts and polyethylene glycol-6000 (PEG-6000) in excised embryo axis of mung bean usingl4C-uracil. Iso-osmotio concentrations of salts and PEG resulted in differential effect on RNA synthesis. PEG inhibited, whereas salt stimulated incorporation of labelled uracil into RNA. Ionic differences on RNA synthesis were very clear as synthesis of RNA was more affected with sulphate than chloride and with sodium than potassium ions. 相似文献
7.
Michio Himeno Yukio Kimura Keizo Hayashiya 《Bioscience, biotechnology, and biochemistry》2013,77(8):1457-1462
Embryos of the silkworm, Bombyx mori L., were dispersed by trypsin and the dissociated cells were cultured for infection with nuclear polyhedrosis virus (NPV) of the silkworm. The monolayer and suspension cultures were infected with NPV. RNA and DNA syntheses in the normal and NPV-infected cells were measured by incorporation of 32P into RNA and DNA fractions. RNA and DNA syntheses in the cells after infection significantly increased over those in control cells (mock infection). The effects of actinomycin D, chloramphenicol and mitomycin C on RNA and DNA syntheses in infected cells were examined. The syntheses were inhibited by the antibiotics. It was suggested that the cellular DNA synthesis was inhibited by the viral infection, because the mitomycin C-resistant DNA synthesis was found in the normal cells but not in the infected cells treated with mitomycin C. The rate of DNA synthesis induced by NPV was immediately dropped to that of control cells by addition of chloramphenicol, while the RNA synthesis induced by NPV was not affected for 6 hr after the addition of chloramphenicol. If the antibiotic did not affect the size of precursor pools, this event suggested that the RNA polymerase concerned with viral RNA synthesis was more stable than the DNA polymerase participating in the viral DNA synthesis. The viral DNA as templates for RNA and DNA syntheses was decomposed by mitomycin C. 相似文献
8.
In cowpea seedlings starch hydrolysing activity increases 35–50 fold on germination for 4 days. This increase in enzyme activity was inhibited by the in vivo addition of 1% glucose but this inhibition was completely overcome by the addition of gibberellic acid (GA3) (10?5 M) and adenosine-3′,5′-cyclic monophosphate (cAMP) (10?5 M). At 5% glucose, GA3 and cAMP were only partially effective. Structural analogues of cAMP failed to relieve the inhibitory effect of glucose. The inhibition by glucose is not direct but RNA and protein synthesis may be involved. Glucose appears to reduce the internal pool of cAMP which causes inhibition of RNA synthesis and decrease in starch hydrolysing activity. Exogenous application of cAMP may replenish the endogenous pool of cyclic nucleotide and thus overcome inhibition of RNA synthesis and enzyme activity. 相似文献
9.
Chromatin- and nuclei-directed ribonucleic Acid synthesis in sugar beet root 总被引:4,自引:3,他引:1
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The synthesis of RNA by chromatin-bound RNA polymerase prepared from sugar beet (Beta vulgaris) root tissue is completely dependent on the presence of a divalent metal (Mg2+ or Mn2+) and the presence of four ribonucleoside triphosphates. Accumulation of labeled acid-insoluble product is inhibited by the addition of RNase and actinomycin D to the reaction. When beet root slices are washed for 25 hours, chromatin-associated RNA polymerase activity increases 7-fold over that of unwashed tissue. This enzyme activity declines with further washing. DNA template availability, as measured by saturating levels of added Escherichia coli RNA polymerase, was also found to follow a pattern similar to that for RNA polymerase. Nearest neighbor frequencies of the RNA synthesized by chromatin isolated from unwashed and washed tissue are different. 相似文献
10.
Purified DNA-dependent RNA polymerases A, B and C isolated from calf thymus contain a significant amount of zinc. Atomic absorption spectroscopy revealed the presence of 6.7, 5.35 and 2.6–4.1 g-atoms of zinc per mole of polymerase A, B and C, respectively. These enzymes are inhibited by treatment with 1,10-phenanthroline at concentrations varying from 10-5 to 10-4 M. However, the addition of zinc ions does not restore fully the activity of 1,10-phenanthroline treated enzymes. Exogenous zinc ions reducein vitro an overall RNA synthesis catalysed by RNA polymerases from calf thymus. In addition to the sites which bind zinc in a specific and stoichiometric way these enzymes possess other classes of binding sites with high and low affinity. Occupancy by exogenous zinc of these additional binding sites inhibits polymerase activity. 相似文献
11.
Characterization of the ribonucleic acid synthesized by isolated rat-liver nuclei 总被引:4,自引:3,他引:1
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1. Isolated rat-liver nuclei incorporated [14C]UMP into RNA when incubated in the presence of Mg2+ and all four ribonucleoside triphosphates. The addition of bentonite to the system diminished the breakdown of the newly synthesized RNA. 2. AMP and CMP were incorporated in the absence of the other added triphosphates, and in the presence of deoxyribonuclease. 3. RNA synthesized in the presence of Mg2+ contained a high proportion of CMP and GMP, and sedimented in the regions of ribosomal RNA and of heavier molecules. About 1% of this RNA hybridized with homologous DNA, and hybrid formation was more effectively inhibited by nuclear RNA than by ribosomal RNA. 4. RNA synthesized in the presence of Mn2+ plus ammonium sulphate had a composition intermediate between that of ribosomal RNA and of DNA, and about 4% of this RNA formed hybrids with DNA. 5. Less than 2% of the newly synthesized RNA was capable of forming ribonuclease- and deoxyribonuclease-resistant complexes. 6. It was concluded that the newly synthesized RNA arose as a result of an asymmetric process and included both ribosomal and DNA-like species. 相似文献
12.
The effect of various drugs on the release of [3H]-serotonin from synaptosomes of reserpine-treated rats was compared with that obtained with synaptosomes of untreated animals.The increase in [3H]-serotonin release induced by d-fenfluramine was virtually abolished by reserpine; the effect of d-norfenfluramine, the main metabolite of fenfluramine, was instead enhanced in synaptosomes of reserpine treated animals. [3H]-serotonin release induced by l-isomers of fenfluramine or norfenfluramine was increased or not affected, respectively, after reserpine treatment.The effects of other drugs, known to activate serotonin mechanisms such as metachlorophenylpiperazine and quipazine, like d-norfenfluramine, were increased by the reserpine treatment.The present data show that [3H]-serotonin can be released by drugs from two pools with different sensitivity to reserpine. The reserpinized synaptosomes could provide useful information on the mechanisms of action of drugs acting on brain serotonin. 相似文献
13.
《International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology》1988,15(6):637-644
The effects of reserpine on the in vivo binding of 3H-Ro 15-1788, (Ro 15-1788: ethyl 8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5a] [1,4]benzodiazepine-3-carboxylate) a selective benzodiazepine antagonist, in the mouse brain were investigated. The biodistributions of tracer amounts of 3H-Ro 15-1788 in mice were significantly altered by pretreatment with reserpine (2.5 or 5.0 mg/kg, 24 h before the tracer administration). The time courses of radioactivity in the brain and the blood following i.v. injection of 3H-Ro 15-1788 with carrier Ro 15-1788 were not changed by pretreatment with reserpine, which suggested that the specific binding process might be altered by reserpine. The degree of alteration in the in vivo binding of 3H-Ro 15-1788 seemed to be dependent upon the dose of reserpine and the duration after the treatment of reserpine. The maximum changes in the biodistribution of 3H-Ro 15-1788 were observed at 1 day after injection of reserpine. The body temperature and the brain monoamine contents (dopamine, norepinephrine and 5-hydroxytryptamine) in mice were measured as indicators of pharmacological effects of reserpine, and good relationships to the degree of changes in the biodistribution of 3H-Ro 15-1788 and either the body temperature or brain monoamine contents, were observed. Furthermore, the changes in the biodistribution of 3H-Ro 15-1788 in the reserpinized mice were significantly suppressed by antidepressant imipramine treatment. These results suggest that it would be possible to detect the in vivo drug interaction with brain benzodiazepine receptors in the living human brain using 11C-Ro 15-1788 and positron emission tomography (PET). 相似文献
14.
The ontogenic development of the transsynaptic induction of adrenal tyrosine hydroxylase (TH), evoked by reserpine and nicotine was studied in control and hypothyroid young rats, aged 3-52 days. The enzymatic induction was measured as an increase in the enzyme activity, since this increase was shown to be impaired either by an inhibitor of RNA synthesis or by a ganglionic blocker. In the control animals, TH induction elicited by reserpine increases between 3 and 32 days of age. In the hypothyroid rats, the enzymatic induction is impaired up to 32 days; at 52 days the induction is similar in both groups of animals. When nicotine is used as a stimulating agent, hypothyroidism still impairs the enzymatic induction at 5 and 21 days, indicating that at least one of the mechanisms inhibited by hypothyroidism is localized in the adrenal chromaffin cells. The present results, taken together with previous findings dealing with adrenal epinephrine secretion, show that the thyroid hormones play a crucial role in the responses of the adrenal medulla to a stimulation in the developing rat, while they have no effect in the adult. 相似文献
15.
Streptovaricins inhibit Focus Formation by MSV (MLV) Complex 总被引:9,自引:0,他引:9
We recently reported that the streptovaricins inhibit the reaction by which DNA is transcribed from the RNA template resident in murine leukaemia virions (MLV)1. The reports2, 3 which first indicated that this DNA polymerase is present in oncogenic RNA viruses have been confirmed and extended4–8. The enzyme provides a mechanism whereby an RNA virus can insert stable genetic information into a host chromosome. Gallo and co-workers described an RNA dependent DNA polymerase in lymphoblastic leukaemic cells which was inhibited by N-demethylrifampicin9 and this antibiotic, together with a number of other rifamycin derivatives, also inhibited the oncogenic viral DNA polymerase10. Like the streptovaricins, the rifamycins are ansa macrolide antibiotics (Fig. 1). 相似文献
16.
James L. White 《Biochemical and biophysical research communications》1978,83(3):1099-1104
3′-deoxyadenosine triphosphate inhibited [3H]UMP incorporation by RNA-dependent RNA polymerases from tobacco and cowpea plants. The inhibition of [3H]UMP incorporation could be reversed by simultaneous addition of higher ATP concentrations but not with increasing concentrations of UTP or when excess ATP was added 10 min after the inhibitor. These results suggest 3′-deoxyadenosine triphosphate competes specifically with ATP in reaction mixtures and results in premature termination of RNA synthesis by RNA-dependent RNA polymerase. 相似文献
17.
Insulin-modulated transport of RNA from isolated live nuclei 总被引:8,自引:0,他引:8
The addition of 3 × 10?7m insulin to a cell-free RNA transport system caused an increase of 50% in the release of messenger-like RNA from 30-min prelabeled rat liver nuclei. Insulin concentrations above 1.2 × 10?6m inhibited RNA release. These hormonal effects were not observed when nuclei were prepared from the insulin-resistant Zucker rat (fa/fa), while the level of stimulation in the heterozygote was approximately one-half that observed with normal liver nuclei. Nuclei prelabeled for 120 min and releasing predominantly ribosomal RNA also did not respond to insulin added to the cell-free system. The hormone appears to affect primarily mRNA transport rather than processing. 相似文献
18.
19.
A fraction containing synaptic vesicles was isolated from rat heart by differential centrifugation, and the uptake of l-[3H]norepinephrine was studied in vitro., Uptake was highly dependent upon time and temperature, and was linear for 6 min at 30° or 4 min at 37°C. About 80% of the measured uptake required both ATP and Mg2+ and was inhibited by nanomolar concentrations of reserpine; no inhibition was obtained with cocaine. These properties are characteristic of storage vesicle uptake as opposed to synaptic membrane uptake. Uptake of norepinephrine was saturable and displayed a single Km value of 2 μM. The uptake was completely stereospecific, as unlabeled dl-norepinephrine was less than half as effective as unlabeled l-norepinephrine in reducing uptake of l-[3H]norepinephrine. Norepinephrine uptake could be inhibited by various phenethylamines and indoleamines following the rank order: reserpine > harmaline > 5-hydroxytryptamine > dopamine > norepinephrine. The vesicle preparation also incorporated [3H]5-hydroxytryptamine and [3H]dopamine. 5-Hydroxytryptamine uptake displayed a Km of 0.5 μM and a maximal uptake equivalent to that seen with norepineph-rine; dopamine uptake followed complex kinetics. Administration of reserpine in vivo or destruction of sympathetic neurons by long-term guanethidine treatment both eliminated the ability of the preparation to take up norepinephrine. Synaptic vesicles of cardiac sympathetic neurons thus resemble vesicles prepared from other central and peripheral catecholaminergic tissues; this method may be used readily to examine drug effects on rat heart synaptic vesicle function. 相似文献
20.
Ribosomal RNA Turnover in Contact Inhibited Cells 总被引:14,自引:0,他引:14
MICHAEL J. WEBER 《Nature: New biology》1972,235(54):58-61
CONTACT inhibition of animal cell growth is accompanied by a decreased rate of incorporation of nucleosides into RNA1–3. Contact inhibited cells, however, transport exogenously-supplied nucleosides more slowly than do rapidly growing cells4,5, suggesting that the rate of incorporation of isotopically labelled precursors into total cellular RNA may be a poor measure of the absolute rate of RNA synthesis by these cells. Recently, Emerson6 determined the actual rates of synthesis of ribosomal RNA (rRNA) and of the rapidly labelled heterogeneous species (HnRNA) by labelling with 3H-adenosine and measuring both the specific activity of the ATP pool and the rate of incorporation of isotope into the various RNA species. He concluded that contact inhibited cells synthesize ribosomal precursor RNA two to four times more slowly than do rapidly growing cells, but that there is little if any reduction in the instantaneous rate of synthesis of HnRNA by the non-growing cells. We have independently reached the same conclusion from simultaneous measurements on the specific radioactivity of the UTP pool and the rate of 3H-uridine incorporation into RNAs (unpublished work of Edlin and myself). However, although synthesis of the 45S precursor to ribosomal RNA is reduced two to four times in contact inhibited cells, the rate of cell multiplication and the rate of rRNA accumulation are reduced ten times. This suggests either “wastage”7 of newly synthesized 45S rRNA precursor, or turnover of ribosomes in contact inhibited cells Two lines of evidence suggest that “wastage” of 45S RNA does not play a significant role in this system. (1) The rate of synthesis of 45S RNA in both growing and contact inhibited cells agrees well with that expected from the observed rates of synthesis of 28S and 18S RNAs (unpublished work of Edlin and myself). Emerson has made similar calculations6. (2) 45S RNA labelled with a 20 min pulse of 3H-uridine is converted in the presence of actinomycin D to 28S and 18S RNAs with the same efficiency (approximately 50%) in both growing and contact inhibited cells. These results indicate that, in order to maintain a balanced complement of ribosomal RNAs, contact inhibited cells must turn over their ribosomes. We present evidence here that rRNA is stable in rapidly growing chick cells, but begins to turn over with a half-life of approximately 35–45 h as cells approach confluence and become contact inhibited. 相似文献