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101.
102.
M. Herrera-Marschitz J. J. Meana W. T. O'Connor M. Goiny M. S. Reid U. Ungerstedt 《Amino acids》1992,2(1-2):157-179
Summary The neuronal origin of extracellular levels of dopamine (DA), acetylcholine (ACh), glutamate (Glu), aspartate (Asp) and gamma-aminobutyric acid (GABA) simultaneously collected from the neostriatum of halothane anaesthetized rats with in vivo microdialysis was studied. The following criteria were applied (1) sensitivity to K+-depolarization; (2) sensitivity to inhibition of synaptic inactivation mechanisms; (3) sensitivity to extracellular Ca2+; (4) neuroanatomical regionality; sensitivity to selective lesions and (5) sensitivity to chemical stimulation of the characterized pathways.It was found that: (1) Extracellular DA levels found in perfusates collected from the neostriatum fulfills all the above criteria and therefore the changes in extracellular DA levels measured with microdialysis reflect actual release from functionally active nerve terminals, and so reflect ongoing synaptic transmission. (2) Changes in neostriatal ACh levels reflect neuronal activity, provided that a ACh-esterase inhibitor is present in the perfusion medium. (3) Extracellular Glu, Asp and GABA could be measured in different perfusion media in the rat neostriatum and probably reflect metabolic as well as synaptic release. However, (4) the majority of the extracellular GABA levels found in perfusates collected from the neostriatum may reflect neuronal release, since GABA levels were increased, in a Ca2+-dependent manner, by K+-depolarization, and could be selectively decreased by an intrinsic neostriatal lesion. (5) It was not possible to clearly distinguish between the neuronal and the metabolic pools of Glu and Asp, since neostriatal Glu and Asp levels were only slightly increased by K+-depolarization, and no changes were seen after decortication. A blocker of Glu re-uptake, DHKA, had to be included in the perfusion medium in order to monitor the effect of K+-depolarization on Glu and Asp levels. Under this condition, it was found (6) that neostriatal Glu and Asp levels were significantly increased by K+-depolarization, although only increases in the Glu levels were sensitive to Ca2+ in the perfusion medium, suggesting that Glu but not Asp is released from vesicular pools. (7) Evidence is provided that selective stimulations of nigral DA cell bodies may lead to changes in release patterns from DA terminals in the ipsilateral neostriatum, which are in turn followed by discrete changes in extracellular levels of GABA and Glu in the same region. Finally, some methodological considerations are presented to clarify the contribution of neuronal release to extracellular levels of amino acid neurotransmitters in the rat neostriatum. 相似文献
103.
Drug distribution studies with microdialysis. III: Extracellular concentration of caffeine in adipose tissue in man 总被引:9,自引:0,他引:9
Microdialysis was applied to estimate the extracellular concentration of caffeine in subcutaneous abdominal adipose tissue of five healthy volunteers after oral administration of approximately 5 mg/kg (300 or 400 mg) of caffeine. The peak extracellular levels were in the range of 20 - 80 microM. The time-course in blood and in extracellular fluid was similar but the plateau concentrations were not closely correlated. The estimated mean concentration of five individuals was similar in blood and extracellular fluid. The intraindividual variation between probes was found to be small compared to the interindividual variation (8% versus 43%). It is concluded that microdialysis yield useful data on drug distribution in man and that distribution to adipose tissue may not strictly follow the concentrations in blood. A comparison with available information of the in vitro properties of caffeine shows that the levels attained in the extracellular fluid were too small to significantly affect phosphodiesterase but sufficiently high to block adenosine receptors. 相似文献
104.
Summary Perinatal asphyxia was induced by keeping pups-containing uterus horns, removed by hysterectomy, in a 37°C or a 30°C water bath. Asphyxia for a period of 21–22 min at 37°C led to a 97% mortality within the first 20 min period following delivery. When the asphyctic period was extended to more than 22 min all the pups died following delivery. When the asphyxia was induced at 30°C, 100% of the delivered pups survived and were accepted by surrogate mothers. The protective effect of hypothermia could be observed even when the pups-containing uterus horns were exposed to a 45–46 min asphyctic period. Pretreatment with dizocilpine (0.2 mg/kg s.c.), or 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(f)quinoxaline (NBQX) (3–30 mg/kg s.c.), administered to the mothers one hour before hysterectomy, reduced slightly the mortality induced by a 21–22 min asphyctic period at 37°C. An increase in survival following a 22–23 min asphyctic period could only be observed after the highest dose of NBQX. 相似文献
105.
Shechinah Felice Choragudi Ganesh Kumar Veeramachaneni BV Raman Bondili JS 《Bioinformation》2014,10(8):507-511
Endo- β-N-acetylgucosaminidases (ENGases) are the enzymes that catalyze both hydrolysis and
transglycosylation reactions. It is of interest to study ENGases because of their ability to synthesize glycopeptides.
Homology models of Human, Arabidopsis thaliana and Sorghum ENGases were developed and their active sites
marked based on information available from Arthrobacter protophormiae (PDB ID: 3FHQ) ENGase. Further, these
models were docked with the natural substrate GlcNAc-Asn and the inhibitor Man3GlcNAc-thiazoline. The catalytic
triad of Asn, Glu and Tyr (N171, E173 and Y205 of bacteria) were found to be conserved across the phyla. The crucial
Y299F mutation showing 3 times higher transglycosylation activity than in wild type Endo-A is known. The hydrolytic
activity remained unchanged in bacteria, while the transglycosylation activity increased. This Y to F change is found
to be naturally evolved and should be attributing higher transglycosylation rates in human and Arabidopsis thaliana
ENGases. Ligand interactions Ligplots revealed the interaction of amino acids with hydrophobic side chains and polar
uncharged side chain amino acids. Thus, structure based molecular model-ligand interactions provide insights into
the catalytic mechanism of ENGases and assist in the rational engineering of ENGases. 相似文献
106.
107.
Elisabeth Ronne-Engström Hans Carlson †Liu Yansheng ‡Urban Ungerstedt §Lars Hillered 《Journal of neurochemistry》1995,65(1):257-262
Abstract: The aim of this study was to evaluate the influence of perfusion media with different glucose concentrations on dialysate levels of lactate, pyruvate, aspartate (Asp), and glutamate (Glu) under basal and hypoxic conditions in rat brain neocortex. Intracerebral microdialysis was performed with the rat under general anesthesia using bilateral probes (o.d. 0.3 mm; membrane length, 2 mm) perfused with artificial CSF containing 0.0 and 3.0 m M glucose, respectively. Basal dialysate levels were obtained 2 h after probe implantation in artificially ventilated animals. Dialysate levels of glucose were also measured for the two different perfusion fluids. The mean absolute extracellular concentration of glucose was estimated by a modification of the no-net-flux method to be 3.3 mmol/L, corresponding to an average in vivo recovery of 6% for glucose. Hypoxia was induced by lowering the inspired oxygen concentration to 3%. Hypoxia caused a disturbance of cortical electrical activity, evidenced by slower frequency and lower amplitudes on the electroencephalogram compared with prehypoxic conditions. This was associated with significant elevations of lactate, Asp, and Glu levels. There were no statistically significant differences in dialysate metabolite levels between the two perfusion fluids, during either normal or hypoxic conditions. We conclude that microdialysis with glucose-free perfusion fluid does not drain brain extracellular glucose in anesthetized rats to the extent that the dialysate lactate, pyruvate, Asp, and Glu levels during basal or hypoxic conditions are altered. 相似文献
108.
Summary The distribution of adrenergic nerves to the male reproductive ducts were studied by applying the highly specific method of Falck and Hillarp for the cellular demonstration of the adrenergic transmitter. In rats, guinea-pigs, and rabbits, the ducts of the testis and the caput epididymidis were found to lack adrenergic varicose terminals, as they also lack cholinesterase positive structures. Adrenergic terminals extend in the vas deferens and epididymis from the prostatic end as far as the upper mid-corpus level of the ductus epididymidis, occurring in the muscle layers and, except in the rat, in the lamina propria. In the cat, ductuli efferentes and caput epididymidis also receive an adrenergic innervation.For skilful technical assistance we are grateful to Mrs. Ulla Flyger and Miss Berith Hansson. The investigation has been supported by a research grant (B 66-257) from the Swedish Medical Research Council.On sabbatical leave from the Biology Department, University of Oregon, Eugene, Oregon, with the aid of a Senior Research Fellowship from the National Council of Child Health and Human Development, National Institutes of Health, U.S. Department of Health, Welfare and Education. The use of facilities of the Department of Physiology I, Karolinska Institutet, with the kind permission of Professor U. S. von Euler and Research Docent R. Eliasson during the course of this study also is greatly appreciated. 相似文献
109.
The regional distribution of various forms of tachykinin-like immunoreactivity (TKLI) was studied in rat brain using radioimmunoassay. TKLI was measured with two different tachykinin-antisera (K12 and E7), which react with neurokinin A (NKA) and neurokinin B (NKB) but not with substance P (SP) and with a specific SP-antiserum. TKLI-K12 and TKLI-E7 were found to have similar regional distributions which were, however, significantly different from that of the substance P-like immunoreactivity (SPLI). Thus, the ratio of the tissue concentrations of TKLI-K12 or TKLI-E7 to that of SPLI was higher in frontal cortex and hippocampus and lower in pons/medulla oblongata than in the other regions studied. Cation-exchange chromatography of neutral water extracts of brain tissue revealed two major immunoreactive components of TKLI-K12 and TKLI-E7, one of which co-eluted with synthetic NKB while the other appeared in the same region as synthetic NKA. The relative quantities of these components varied depending on the brain region studied. No TKLI-K12 or TKLI-E7 co-eluted with synthetic SP. Almost all of the SPLI in acetic acid or water extracts of brain tissue eluted as a single chromatographic component in the same position as synthetic SP. Potassium-stimulated in vivo release of TKLI-K12, TKLI-E7 and SPLI in striatum of rat brain could be demonstrated using intracerebral dialysis. The present results imply that tachykinins, which may serve as neurotransmitters or neuromodulators, are present in different proportions in different regions of rat brain. 相似文献