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71.
Abstract: We have studied the regional distribution and characteristics of polyamine-sensitive [3H]ifenprodil binding sites by quantitative autoradiography in the rat brain. In forebrain areas ifenprodil displaced [3H]ifenprodil (40 nM) in a biphasic manner with IC50 values ranging from 42 to 352 nM and 401 to 974 µM. In hindbrain regions, including the cerebellum, ifenprodil displacement curves were monophasic with IC50 values in the high micromolar range. Wiping studies using forebrain slices (containing both high- and low-affinity sites) or cerebellar slices (containing only the low-affinity site) showed that high- and low-affinity ifenprodil sites are sensitive to spermine and spermidine, to the aminoglycoside antibiotics neomycin, gentamicin, and kanamycin, and to zinc. Two calmodulin antagonists, W7 and calmidazolium, also displaced [3H]ifenprodil from both sites. Other calmodulin antagonists, including trifluoperazine, prenylamine, and chlorpromazine, selectively displaced [3H]ifenprodil from its low-affinity site in hindbrain and forebrain regions. High-affinity [3H]ifenprodil sites, defined either by ifenprodil displacement curves or by [3H]ifenprodil binding in the presence of 1 mM trifluoperazine, were concentrated in the cortex, hippocampus, striatum, and thalamus with little or no labeling of hindbrain or cerebellar regions. This distribution matches that of NMDAR2B mRNA, supporting data showing that ifenprodil has a preferential action at NMDA receptors containing this subunit. Low-affinity [3H]ifenprodil sites have a more ubiquitous distribution but are especially concentrated in the molecular layer of the cerebellum. [3H]Ifenprodil was found to bind to calmodulin-agarose with very low affinity (IC50 of ifenprodil = 516 µM). This binding was displaced by calmodulin antagonists and by polyamines, with a potency that matched their displacement of [3H]ifenprodil from its low-affinity site in brain sections. However, the localization of the low-affinity [3H]ifenprodil site does not strictly correspond to that of calmodulin, and its identity remains to be further characterized. The restricted localization of high-affinity [3H]ifenprodil binding sites to regions rich in NMDAR2B subunit mRNA may explain the atypical nature of this NMDA antagonist. 相似文献
72.
Chronic Nicotine Administration Differentially Affects Neurotransmitter Release from Rat Striatal Slices 总被引:6,自引:3,他引:3
Abstract: The objective of these experiments was to determine whether the chronic administration of nicotine, at a dose regimen that increases the density of nicotine binding sites, alters the nicotine-induced release of [3H]dopamine ([3H]DA), [3H]norepinephrine ([3H]NE), [3H]serotonin ([3H]5-HT), or [3H]acetylcholine ([3H]ACh) from rat striatal slices. For these experiments, rats received subcutaneous injections of either saline or nicotine bitartrate [1.76 mg (3.6 µmol)/kg, dissolved in saline] twice daily for 10 days, and neurotransmitter release was measured following preloading of the tissues with [3H]DA, [3H]NE, [3H]5-HT, or [3H]choline. Chronic nicotine administration did not affect the accumulation of tritium by striatal slices, the basal release of radioactivity, or the 25 mM KCl-evoked release of neurotransmitter. Superfusion of striatal slices with 1, 10, and 100 µM nicotine increased [3H]DA release in a concentration-dependent manner, and release from slices from nicotine-injected animals was significantly (p < 0.05) greater than release from saline-injected controls; release from the former increased to 132, 191, and 172% of release from the controls following superfusion with 1, 10, and 100 µM nicotine, respectively. Similarly, [3H]5-HT release increased in a concentration-related manner following superfusion with nicotine, and release from slices from nicotine-injected rats was significantly (p < 0.05) greater than that from controls. [3H]5-HT release from slices from nicotine-injected rats evoked by superfusion with 1 and 10 µM nicotine increased to 453 and 217%, respectively, of release from slices from saline-injected animals. The nicotine-induced release of [3H]NE from striatal slices was also concentration dependent but was unaffected by chronic nicotine administration. [3H]ACh release from striatal slices could not be detected when samples were superfused with nicotine but was measurable when tissues were incubated with nicotine. The release of [3H]ACh from slices from nicotine-injected rats was significantly (p < 0.05) less than release from controls and decreased to 36, 83, and 77% of control values following incubation with 1, 10, or 100 µM nicotine, respectively. This decreased [3H]ACh release could not be attributed to methodological differences because slices from nicotine-injected rats incubated with nicotine exhibited an increased [3H]DA release, similar to results from superfusion studies. In addition, it is unlikely that the decreased release of [3H]ACh from striatal slices from nicotine-injected rats was secondary to increased DA release because [3H]ACh release from slices from hippocampus, which is not tonically inhibited by DA, also decreased significantly (p < 0.05) in response to nicotine; hippocampal slices from nicotine-injected rats incubated with 1 and 10 µM nicotine decreased to 42 and 70%, respectively, of release from slices from saline-injected animals. Results indicate that the chronic administration of nicotine increases the ability of nicotine to induce the release of [3H]DA and [3H]5-HT and decreases the ability of nicotine to evoke the release of [3H]ACh but does not alter the nicotine-induced release of [3H]NE from brain slices. 相似文献
73.
Abstract: Solubilization of rat striatal membranes with sodium cholate, followed by reconstitution into phospholipid vesicles, leads to a 6.5-fold increase in the agonist high-affinity binding sites of the D1 dopamine receptor. These high-affinity binding sites display differential sensitivity toward temperature. When reconstituted receptors were preincubated for 1 h at 0–4°C (on ice) or at 22°C (room temperature) followed by radioligand binding assays with dopamine, neither the high-affinity values of the receptor for dopamine nor the percent receptors in the high-affinity state (31–39%) were changed from control reconstituted receptors, which were not subject to any preincubations. At 30°C, there was a partial loss in the number of high-affinity D1 receptors with only 25% of the total receptor population in the high-affinity state; there was no change in the affinity values of the high-affinity binding sites. At 37°C, there was a 40% loss in total number of D1 receptor binding sites. All the high-affinity binding sites were lost and the remaining 60% of binding activity represented the low-affinity binding state of the receptor. These results indicate that the high-affinity binding sites of the reconstituted D1 dopamine receptors are uniquely sensitive to higher temperatures. 相似文献
74.
S. Chao C. Baysdorfer O. Heredia-Diaz T. Musket G. Xu E. H. Coe Jr 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,88(6-7):717-721
We report here the results of mapping a set of 92 leaf cDNA clones in maize. The ends of each of these cDNA clones have previously been partially sequenced, and the sequence comparison has revealed the putative function for 28 clones. It is expected that the RFLP map developed using these expressed sequence tags will be of great importance for future maize genome analysis, such as for PCR-based gene mapping or gene function identification.Contribution from the Missouri Agricultural Experiment Station. Journal Series N. 12,019. 相似文献
75.
KENNETH C. EMBERTON 《Biological journal of the Linnean Society. Linnean Society of London》1994,53(2):175-187
Nine species of Madagascan acavid land snails were compared in a phylogenetic context. The two most plesiomorphic, Clauator johnsoni and C. moreleti, differ from the others by their high-spired shells, short tentacles, short tails, long necks, and crawling mode of hitching the shell along the ground. In the seven more apomorphic species, the crawling mode is smooth, with the shell resting on the tail, and the relative lengths of tail and shell correlate significantly. Among these seven species, three pairs of closest relatives (Helicophanta petiti and H. uesicalis, H. farafanga and H. souuerbiana, Ampeltta decaryi and A. julii) show evidence of phylogenetic constraints on ranked shell size. Aestivation site (as tentatively inferred from rare data) does not correlate with shell shape or size: burrowers have H/D = 2.7 to 0.6 and D = 70 to 25 mm; arboreals have H/D = 0.8 to 0.5 and D = 70 to 30 mm; the species with both the highest spire and the smallest diameter (C. moreleti) is neither a burrower or an arboreal, but stays on the ground surface. Inferred aestivation sites are randomly distributed phylogenetically. Climate shows no correlation, except that the arborcals are only from humid to wet regimes. Uniform shell colouration occurs only in burrowers (C. johnsoni, H. petiti, H. uestcalis), but disruptive shell colouration occurs in all others, including burrowers (H. farafanga, A. decaryt), ground-surface aestivators (C. moreleti), arboreals (H. souuerbiana, A. julii), and semi-arboreals (Ampefita subfunebris). Among all nine species, burrowers have significantly thicker shells (than their close relatives of similar size), wider bodies, and longer snouts than non-burrowers (H. souuerbiana is exceptional in being arboreal despite its huge size and in having the broad foot and snout of a burrower). Thus, although there is some evidence for phylogenetic constraints, natural selection for aestivation and crawling behaviours seems to have dominated the evolution of external body morphology and of shell thickness (but not shell size and shape) in these snails. 相似文献
76.
Serafina Salvati Lucilla Attorri Lorenzo Malvezzi Campeggi Antonella Olivieri Mariella Sorcini Stefano Fortuna Annita Pintor 《Neurochemical research》1994,19(9):1181-1186
The effect of hypothyroidism on the lipid composition of synaptosomes, density and affinity of muscarinic receptor sites, and acetylcholinesterase activity in the cerebral cortex of young and aged rats was investigated. The animals were made hypothyroid by adding 0.05% propyl-2-thiouracil to their drinking water for four weeks. This pathological state induced an increase in the relative percentage of sphingomyelin in young rats. In aged rats hypothyroidism induced a decrease of sphingomyelin and glycerophosphocholine and an increase of cholesterol. The effect of hypothyroid state on cerebral cortex resulted in an increase of acethylcholinesterase activity both in young and aged rats and was also reflected in an increase of density of M1-AChRs but only in the former. 相似文献
77.
78.
István Papp László Dorgai Péter Papp Erzsébet Jónás Ferenc Olasz László Orosz 《Molecular & general genetics : MGG》1993,240(2):258-264
Bacteriophage 16-3 inserts its genome into the chromosome of Rhizobium meliloti strain 41 (Rm41) by site-specific recombination. The DNA regions around the bacterial attachment site (attB) and one of the hybrid attachment sites bordering the integrated prophage (attL) were cloned and their nucleotide sequences determined. We demonstrated that the 51 by region, where the phage and bacterial DNA sequences are identical, is active as a target site for phage integration. Furthermore it overlaps the 3 end of a putative proline tRNA gene. This gene shows 79% similartiy to the corresponding proline tRNA-like genomic target sequence of certain integrative plasmids in Actinomycetes. 相似文献
79.
Fragment X (LMrFX) was obtained as low molecular weight preparations from a late stage 2 plasmin digest of human fibrinogen. The thrombin-treated LMrFX preparations, which resulted in impaired polymerization, were further subfractionated into polymerized and non-polymerized components. The fractions were examined by SDS-PAGE and immunochemical methods. In polymerized fractions, more peptide bands were observed on SDS-PAGE in the reduced state than in non-polymerized fractions. Both fractions contained a similar number of internal cleavages in the A, Bβ and γ chains, which are linked by disulfide bonds. Thus, the partial deficiencies in polymerization sites of the carboxy terminal region of the γ chain and the amino terminal portions of the Bβ chain, as well as internal cleavage, were considered to participate in the impairment of the thrombin-induced polymerization of LMrFX. 相似文献
80.
David H. Rosenblatt John F. Manning Jr Carlo D. Montemagno 《Soil & Sediment Contamination》1994,3(1):1-27
Plans are being formulated for in situ bioremediation of a subsurface plume of diesel fuel No. 2 that resulted from an accidental fuel release. Raoult's law and the aqueous solubilities of the toxic components were used to estimate organic contaminant concentrations in leachate from the untreated fuel mass. Carcinogenic risks and noncarcinogenic hazard indices were calculated for undiluted leachate. An 80% decrease in hydrocarbon mass and increases in the average molecular weights of the component fractions were assumed to result from the treatment. Sample calculations are provided to show how to evaluate results of analyses for petroleum hydrocarbons after bioremediation. 相似文献