共查询到20条相似文献,搜索用时 15 毫秒
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Preparation and properties of a cell-free, hormonally responsive adenylate cyclase from guinea pig brain 总被引:7,自引:5,他引:2
—The accumulation of cyclic adenosine 3′,5′-monophosphate (cyclic AMP) was studied in cell-free homogenates of guinea pig brain. Homogenates, prepared in Krebs-Ringer buffer, responded markedly to the addition of neurohormones with an increased rate of cyclic AMP synthesis; preparations from cerebellum, cerebral cortex, and hippocampus responded to a degree approximating that achieved with slices of these areas of guinea pig brain. Adenylatc cyclase activity was seen only when cyclic AMP was measured by a [3H]adenine prelabelling technique or when total cyclic AMP was measured by radioimmunoassay; [32P]ATP did not serve as a substrate for this preparation of the enzyme. The adenylate cyclase was paniculate and required a Krebs Ringer buffer; use of tris, or tris with Mg2+ and Ca2+, resulted in a preparation totally devoid of hormonal stimulation. Digestion by purified beef heart cyclic nucleotide phosphodiesterase, Dowex chromatography, solubility in Ba(OH)2-ZnSO4 mixtures, and two thin layer chromatographic systems demonstrated that the product of the hormonally stimulated adenylate cyclase preparation was cyclic AMP. The selectivity of hormonal stimulation and the adrenergic character of the hormonal receptors from different brain areas were maintained in the cell-free preparation. However, simultaneous stimulation with two different neurohormones resulted in additive responses, rather than in the potentiation observed in preparations of slices of brain. 相似文献
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Enhanced serotonin-stimulated adenylate cyclase activity in membranes from adult guinea pig hippocampus 总被引:4,自引:0,他引:4
We have developed an assay for serotonin (5-HT) stimulation of adenylate cyclase activity in membranes from adult guinea pig hippocampus. The response to 5-HT is concentration-dependent, with an EC50 of 0.01 microM, a shallow slope, and mean maximal stimulation of 90% over basal activity. The response to 5-HT is GTP-dependent and additive to the maximal stimulation by histamine. Micromolar concentrations of the known 5-HT receptor agonists, tryptamine and 5-methoxytryptamine, also stimulate cAMP production in this system, and their effect is not additive to that elicited by a maximal concentration of 5-HT. These results are consistent with the hypothesis that the response to 5-HT is elicited through a distinct receptor coupled to adenylate cyclase; the magnitude and the reproducibility of the 5-HT response in this system should make it useful for receptor classification. To examine the effect of prior exposure to endogenous 5-HT on the responsiveness of the system, we assayed 5-HT stimulation of enzyme activity in membranes prepared from animals 25-27 hrs after treatment with a single injection of reserpine (5 mg/kg, i.p.). The mean maximal stimulation of adenylate cyclase by 5-HT was increased to 150% over basal activity with no effect on the EC50 or slope of the 5-HT concentration-response curve. Reserpine pretreatment did not affect basal activity or histamine-stimulated adenylate cyclase activity. These results are discussed in the context of a hypothesis that endogenous 5-HT normally exerts a desensitizing effect on its receptors in situ. 相似文献
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Demonstration of adenylate cyclase coupled adenosine receptors in guinea pig ventricular membranes 总被引:1,自引:0,他引:1
E Leung C I Johnston E A Woodcock 《Biochemical and biophysical research communications》1983,110(1):208-215
Adenylate cyclase in homogenates of guinea pig ventricles was inhibited by the stable adenosine analogs N6-phenylisopropyladenosine (PIA) and 5(1)-N-ethylcarboxamidoadenosine (NECA). Inhibition required GTP and was enhanced by sodium ion. The maximum inhibition observed was 35.1 +/- 1.1%, the EC50 (95% confidence limits, n) for PIA and NECA were 0.20 microM (0.17-0.25 microM, 6) and 0.66 microM (0.26-1.7 microM, 4) respectively. 8-Phenyltheophylline (10 and 100 microM) and isobutylemethylxanthine (100 microM) antagonized the inhibitory effects of the adenosine analogs. These results indicate that adenosine receptors of the inhibitory type (RI or A1) are present in guinea pig myocardium and may mediate some of the cardiac responses to adenosine. 相似文献
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Ca2+-dependent regulation of guinea pig brain adenylate cyclase 总被引:8,自引:0,他引:8
M T Piascik P L Wisler C L Johnson J D Potter 《The Journal of biological chemistry》1980,255(9):4176-4181
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M Hitchcock 《Journal of immunology (Baltimore, Md. : 1950)》1977,118(2):578-583
Cyclic AMP has been implicated in the regulation of the immunologic release of histamine from lung and other tissues and cell types. The mechanism whereby intracellular levels of cAMP are altered during mediator release was investigated. Measurements of histamine, adenylate cyclase, and cAMP phosphodiesterase activities were made in actively and passively sensitized guinea pig lung after challenge with antigen. A transient decrease in basal adenylate cyclase activity occurred which returned to control levels after histamine release. There was no change in cAMP phosphodiesterase activity determined at substrate concentrations of 1 mM and 0.01 mM. The adenylate cyclase response did not occur under the following conditions: 1) incubation of nonsensitized lung with antigen, 2) incubation of sensitized lung with antigen in the absence of extracellular calcium, and 3) incubation of nonsensitized lung with compound 48/80. These observations indicate 1) the adenylate cyclase response and the immunologic release of histamine are intimately related, and 2) the reduction in intracellular levels of cAMP which have been reported to occur during immunologic histamine release are mediated via adenylate cyclase. 相似文献
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Creutzfeldt-Jakob infection increases adenylate cyclase activity in specific regions of guinea pig brain 总被引:3,自引:0,他引:3
Creutzfeldt-Jakob disease is a slow, infectious, progressive neurological disorder which results in human dementia. Synaptic membranes from various brain regions of guinea pigs infected with Creutzfeldt-Jakob disease show increased guanyl nucleotide- or 5-hydroxytryptamine-mediated activation of adenylate cyclase. This increased enzyme activity appears due, primarily, to facilitated 'coupling' between the GTP-binding protein which stimulates adenylate cyclase (GNs) and the catalytic moiety of that enzyme rather than increased sensitivity to 5-hydroxytryptamine. It is possible that this phenomenon is due to direct effects of the Creutzfeldt-Jakob infectious agent, or a pathological product resulting from that agent, upon synaptic membrane adenylate cyclase. 相似文献
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Dissociation between adenosine receptors and adenylate cyclase in the smooth muscle of guinea pig myometrium 总被引:2,自引:0,他引:2
We have previously demonstrated that adenosine causes contraction of guinea-pig myometrium in a fashion consistent with the presence of a purinergic receptor of the A1 subtype. Incubation of guinea-pig uterine smooth muscle membranes with the stable adenosine analogue [3H]cyclohexyladenosine [( 3H]CHA) resulted in rapid, reversible association of radioligand to saturable sites. The affinity (KD) of the receptor for [3H]CHA determined from kinetic experiments (3.14 nM) is in good agreement with that determined in saturation experiments (KD = 4.5 nM). Scatchard analysis of specific [3H]CHA binding (Bmax = 79 fmol/mg protein) is consistent with a single class of binding sites for [3H]CHA. Computer analysis of competition of [3H]CHA binding by the stereoisomers of phenylisopropyl adenosine, R-PIA (KI = 5.3 nM) and S-PIA (KI = 69 nM), as well as the 5'-substituted analogue, ethylcarboxamide adenosine (NECA; KI = 4.2 nM) suggest that [3H]CHA binding occurs to a single class of receptors of the AI subtype. Contractile studies employing these agents reveal that the relative order of potency, based on ED50 values, correlates well with the relative order of competition of agonist binding, based on equilibrium binding constants. Direct assay of myometrial adenylate cyclase failed to show that adenosine receptors in this smooth muscle are coupled to adenylate cyclase. We conclude here that a smooth muscle adenosine receptor is not coupled to adenylate cyclase, yet subserves muscle contraction. These data are important in light of recent attempts to classify adenosine receptors as dual regulators of adenylate cyclase. 相似文献
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Vesicles from guinea pig cerebral cortex prepared by homogenization in Krebs-Ringer buffer contained adenylate cyclase activity which was stimulated by the acidic amino acids, cysteine sulfinic and glutamic acids, and by norepinephrine as well as by an alkaloid, veratridine. With these vesicular preparations the concentrations of amino acids required for half-maximal stimulation were about 30 muM, only about 1/30 those necessary with intact-cell preparations. Nearly additive effects were observed when either of the active amino acids was combined with norepinephrine at their optimal concentrations. 相似文献
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H. H. Vink 《Antonie van Leeuwenhoek》1955,21(1):446-448
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L E Aseeva L A Shevchenko N Ia Shimaniuk B D Rublev B N Mishan'kin 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》1987,(7):59-63
The biological action of Y. pestis adenylate cyclase on peritoneal leukocytes of guinea pigs has been studied by means of chemiluminescence. Y. pestis adenylate cyclase is supposed to contribute to the "oxidation" explosion of phagocytes in plague. 相似文献
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Y L Lai W J Lamm D L Luchtel J Hildebrandt 《Journal of applied physiology (Bethesda, Md. : 1985)》1984,56(2):308-314
A special phenomenon (difficult to inflate and deflate) occurring in the postmortem guinea pig lungs was studied in 40 animals. Thirty minutes after excision of the lungs or exsanguination, less than 50% of the lungs could be inflated even at high inflation pressure (34 cmH2O), and most gas was trapped during deflation. The amount of trapped gas volume at 30 min was related to the degree of lung inflation maintained during the 5- to 30-min period after exsanguination. Since stiffness of the lung tissue was unlikely to explain the phenomenon, we speculated airway obstruction as the major factor. No foam or bubbles were found in larger airways and we thus hypothesized that the obstruction was due to bronchoconstriction. This was confirmed histologically in that the lumina of both bronchi and bronchioles were constricted. The latent period to the onset of this constriction was short (approximately 5 min). It was not associated with O2 availability but was delayed an additional 15 min by a thromboxane inhibitor (dazoxiben). Neither maintaining lung temperature at 37 degrees C nor vagotomy and/or cervical transection prevented the constriction. Without exsanguination, onset of bronchoconstriction was delayed by about 1 h. We conclude that postmortem bronchoconstriction may be caused by release of an endogenous constrictor agent. 相似文献
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Massive postmortem bronchoconstriction in guinea pig lungs 总被引:1,自引:0,他引:1
Lai Y. L.; Lamm W. J.; Luchtel D. L.; Hildebrandt J. 《Journal of applied physiology》1984,56(2):308-314
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In thoroughly washed guinea pig fundic gastric mucosal membranes, NaCl markedly potentiates the maximal histamine-stimulated adenylate cyclase activity and increases the concentration of histamine required for half-maximal effect (EC50). The apparent dissociation constants for the antagonists cimetidine and metiamide are only slightly increased in the presence of NaCl. Potassium chloride does not change the histamine EC50 but does increase the maximal histamine-stimulated adenylate cyclase activity. These results suggest that Na and K ions may play an important role in the regulation of histamine-sensitive adenylate cyclase in gastric mucosa. The effect of the Na ion appears to be more specific for histamine H2 receptor agonists than for antagonists. 相似文献
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