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1.
IgM and IgD genes of the Japanese flounder were cloned and characterized from a genomic bacterial artificial chromosome (BAC) library. The gene contained four constant region exons (C1–C4), and two transmembrane exons (TM1 and TM2), which conforms to the organizational pattern of all other vertebrate -chain genes examined so far. In the same BAC clone, the gene, which is homologous to the IgD gene in mammals and teleost fish, was found immediately (0.9 kb) downstream of the IgM gene. This gene encoded seven exons (C1–C7) and two TM exons (TM1 and TM2) and had no duplication of C1-C2, as found in Atlantic cod, or C2-C3-C4, as found in Atlantic salmon and channel catfish. Phylogenetic and sequence analyses strongly suggest that teleost is more closely related to non-IgM isotypes than IgM isotypes. The heavy chain (IgH) locus of Japanese flounder, which encodes m, s and m, was found to be fully functional.  相似文献   

2.
The effect of interspecific competition and element additions (N and P) on four grassland species (Poa pratensis, Lolium perenne, Festuca valida, Taraxacum officinale) grown under field conditions was studied. Two grasses (L. perenne, F. valida) grown in monoculture (absence of competition) showed lower carbon isotope discrimination (13C) and enriched 15N values. Nitrogen addition (as urea) had inconsistent effects on species 13C while caused enrichment of 15N of P. pratensis and F. valida but strong depletion of 15N of T. officinale. Phosphorous had no significant effect on 13C but depleted 15N of all species.  相似文献   

3.
T-cell clones expressing the T-cell receptor (Tcr) were generated from peripheral blood lymphocytes (PBLs) and from a thymus sample. In the panel of ten thymus-derived clones, four Tcr phenotypes [as defined by the reaction of monoclonal antibodies (mAbs) directed against known V and V regions] were identified. All the clones lacked expression of the V3 V region, while seven clones were V1+ . V1 was found in combination with V9 or with undefined VVregions. In addition, two other Tcr phenotypes were identified on these clones: V9+ V1 V3 and V9 V1 V3 One of the clones expressed CD4 and another was CD8positive. The remaining clones were CD4 CD8. In the panel of 76 PBL-derived, Tcr-bearing clones, five Tcr phenotypes could be identified. In contrast to the thymus-derived clones, 30% of the clones were V3+ whereas V1 was expressed by a minority of the clones only. One clone was CD4-positive and approximately 30% of the clones were CD8-positive. Four of the five mAb-defined Tcr phenotypes could be identified on both thymus and PBL-derived T-cell clones. However, biochemical analysis of the Tcrs demonstrates differences in the usage of Ct- and C2-encoded y chains by T cells derived from the thymus and PBLs. The results therefore indicate that, at the clonal level, similarities and differences exist between the Tcr repertoires expressed in the thymus and by PBLs. Furthermore, they indicate that combinatorial Tcr heterogeneity is larger than has so far been described. The receptor diversity, combined with the potential of Tcr+ cells to express CD4 or CD8, indicates that these cells are a heterogeneous population that might mediate a number of immune functions.  相似文献   

4.
Summary Two double heterozygous 0/0 thalassemic sibs of Mexican descent were studied. The father had a 0/0 genotype, while the mother, one sib and several maternal relatives were 0/0 heterozygotes. Parental consanguinity and an apparently low frequency of thalassemia among Mexicans suggested a possible common origin of both 0 and 0 genes. A hypothesis to explain such a possibility is proposed on the basis of a partial mispairing between 0 and genes followed by a crossing-over which would results in a 0 recombinant gene. This hypothesis could also be extended to explain the 22 gluala, 22 alaglu and 116 arghis Hb variants as recombinants from double crossing-over between and mispaired genes for which the name interstitial-Lepore is proposed.  相似文献   

5.
Mild cerebral anoxic/ischemic/stress insults promote tolerance and thereby protect the brain from subsequent lethal anoxic/ischemic insults. We examined whether specific activation of PKC , , , or isoforms is associated with ischemic preconditioning (IPC) in rat brain. IPC was produced by a 2-minute global cerebral ischemia. Membrane and cytosolic fractions of the hippocampi were immunoblotted using specific antibodies for PKC, , , and . PKC showed a significant translocation to the membrane fraction from 30 min to 4 h and PKC at 4 h following IPC. In contrast, the membrane/cytosol ratio of PKC showed a tendency to decrease at 30 min and 8 h, and the membrane/cytosol ratio of PKC was significantly decreased from 30 min to 24 h following IPC. These findings indicate PKC isoform-specific membrane translocations in the hippocampus after brief global brain ischemia and suggest that activation of PKC and PKC may be associated with IPC-induced tolerance in the rat hippocampus.  相似文献   

6.
Summary The 13C values of submerged aquatic plants from contrasting but relatively defined habitats, and the 13C values of emergent, floating and submerged leaves of dimorphic aquatic plants, were measured. In many instances the 13C values of dissolved inorganic carbon in the water were also measured. Plant 13C values in the vicinity of-40 to-50 were found in rapidly flowing spring waters with carbonate 13C values of-16 to-21, consistent with the notion that species such as Fontinalis antipyretica almost exclusively assimilate free CO2 via RuP2 carboxylase. Plant 13C values in the vicinity of-10 to-15 in sluggish water with carbonate 13C values of about-5 were observed, consistent with the notion that boundary layer diffusion and/or HCO3 - uptake may determine the 13C value of submerged aquatic plants in these circumstances. Comparisons of 13C values of the same or related species growing in waters of similar carbonate 13C value but different flow rates confirmed this view; more negative 13C values were frequently associated with plants in fast moving water. In Britain, but not in Finland, the 13C values of submerged leaves of dimorphic plants were almost invariably more negative than in aerial leaves. The 13C value of carbonate from chalk streams and in acid springs indicate substantial inputs of respiratory CO2, as opposed to atmospheric carbon. The contributions of these variations in 13C of the carbon source, and of isotope fractionation in diffusion, to the 13C value of submerged parts of dimorphic plants is discussed.  相似文献   

7.
We have previously shown that mitochondrial membrane potential () drop promoted by prooxidants and Ca2+ can be reversed but not sustained by ethylene glycol-bis(-aminoethylether)-N,N,N,N-tetraacetic acid (EGTA) unless dithiothreitol (DTT), a disulfide reductant, is also added [Valle, V. G. R., Fagian, M. M., Parentoni, L. S., Meinicke, A. R., and Vercesi, A. E. (1993).Arch. Biochem. Biophys. 307, 1–7]. In this study we show that catalase or ADP are also able to potentiate this EGTA effect. When EGTA is added long after (12 min) the completion of swelling or elimination, no membrane resealing occurs unless the EGTA addition was preceded by the inclusion of DTT, ADP, or catalase soon after was collapsed. Total recovery by EGTA is obtained only in the presence of ADP. The sensitivity of the ADP effect to carboxyatractyloside strongly supports the involvement of the ADP/ATP carrier in this mechanism. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of solubilized membrane proteins shows that protein aggregation due to thiol cross-linkage formed during drop continues even after is already eliminated. Titration with 5,5-dithio-bis(2-nitrobenzoic acid) supports the data indicating that the formation of protein aggregates is paralleled by a decrease in the content of membrane protein thiols. Since the presence of ADP and EGTA prevents the progress of protein aggregation, we conclude that this process is responsible for both increased permeability to larger molecules and the irreversibility of drop. The protective effect of catalase suggests that the continuous production of protein thiol cross-linking is mediated by mitochondrial generated reactive oxygen species.  相似文献   

8.
Here we report on the use of iodination of the membrane-bound nicotinic acetylcholine receptor (nAChR) from Torpedo californica electric tissue in order to define surface-exposed portions of the receptor molecule. Membrane-bound nAChR was 125I-iodinated using the oxidation agent Iodo-Gen. The iodinated subunits were separated by preparative gel electrophoresis, desalted, and cleaved with trypsin. The resulting peptides were separated by reverse-phase HPLC and the radioactive peptides were identified by mass spectrometry and protein sequencing. For the -subunit, we identified five iodinated peptides containing the tyrosine residues Tyr17, Tyr74, Tyr365, Tyr372, and Tyr428. The surface exposition of these amino acids is in agreement with the four-transmembrane-segment model (4TM model) of the nAChR, but the assignment to the intra- or extracellular surface is doubtful. According to this model, the N-terminal portion of the receptor subunits including the iodinated residues Tyr17 and Tyr74 is extracellular and Tyr372 as a site of tyrosine phosphorylation is located on the cytoplasmic side. But since this latter residue is among the first to be iodinated using an immobilized iodination agent, its true position with respect to the membrane bilayer is not clear.  相似文献   

9.
Summary Carbon isotope composition, photosynthetic gas exchange, and nitrogen content were measured in leaves of three varieties of Metrosideros polymorpha growing in sites presenting a variety of precipitation, temperature and edaphic regimes. The eight populations studied could be divided into two groups on the basis of their mean foliar 13C values, one group consisting of three populations with mean 13C values ca.-26 and another group with 13C values ca.-28. Less negative 13C values appeared to be associated with reduced physiological availability of soil moisture resulting from hypoxic conditions at a poorly drained high elevation bog site and from low precipitation at a welldrained, low elevation leeward site. Gas exchange measurements indicated that foliar 13C and intrinsic wateruse efficiency were positively correlated. Maximum photosynthetic rates were nearly constant while maximum stomatal conductance varied substantially in individuals with foliar 13C ranging from-29 to-24. In contrast with the patterns of 13C observed, leaf nitrogen content appeared to be genetically determined and independent of site characteristics. Photosynthetic nitrogenuse efficiency was nearly constant over the range of 13C observed, suggesting that a compromise between intrinsic water- and N-use efficiency did not occur. In one population variations in foliar 13C and gas exchange with leaf cohort age, caused the ratio of intercellular to atmospheric partial pressure of CO2 predicted from gas exchange and that calculated from 13C to be in close agreement only in the two youngest cohorts of fully expanded leaves. The results indicated that with suitable precautions concerning measurement protocol, foliar 13C and gas exchange measurements were reliable indicators of potential resource use efficiency by M. polymorpha along environmental gradients.  相似文献   

10.
The feasibility of using nitrogen and oxygenisotope ratios of nitrate (NO3 ) forelucidating sources and transformations ofriverine nitrate was evaluated in a comparativestudy of 16 watersheds in the northeastern U.S.A. Stream water was sampled repeatedly at theoutlets of the watersheds between January andDecember 1999 for determining concentrations,15N values, and 18Ovalues of riverine nitrate.In conjunction with information about land useand nitrogen fluxes,15Nnitrate and18Onitrate values providedmainly information about sources of riverinenitrate. In predominantly forested watersheds,riverine nitrate had mean concentrations ofless than 0.4 mg NO3 -N L–1,15Nnitrate values of lessthan +5, and 18Onitratevalues between +12 and +19. This indicatesthat riverine nitrate was almost exclusivelyderived from soil nitrification processes withpotentially minor nitrate contributions fromatmospheric deposition in some catchments. Inwatersheds with significant agricultural andurban land use, concentrations of riverinenitrate were as high as 2.6 mg NO3 -NL–1 with 15Nnitratevalues between +5 and +8 and18Onitrate values generallybelow +15. Correlations between nitrateconcentrations, 15Nnitratevalues, and N fluxes suggest that nitrate inwaste water constituted a major, and nitrate inmanure a minor additional source of riverinenitrate. Atmospheric nitrate deposition ornitrate-containing fertilizers were not asignificant source of riverine nitrate inwatersheds with significant agricultural andurban land use. Although complementary studiesindicate that in-stream denitrification wassignificant in all rivers, the isotopiccomposition of riverine nitrate sampled at theoutlet of the 16 watersheds did not provideevidence for denitrification in the form ofelevated 15Nnitrate and18Onitrate values. Relativelylow isotopic enrichment factors for nitrogenand oxygen during in-stream denitrification andcontinuous admixture of nitrate from theabove-described sources are thought to beresponsible for this finding.  相似文献   

11.
A combination of stable isotope studies and 14Cdating were used to identify the main sources andprocesses controlling streamwater DOC and TIC in atemperate non-forested watershed. 13Cvalues for terrestrial (–24.9 to –29.1) and aquatic(–30.5 to –33.5) plants were similar to valuesreported in the literature for similar ecosystems.13C values for DOC in soil solution andstreamwater were consistent with soil and terrestrialvegetation, indicating that the terrestrial ecosystemis the dominant source of aquatic DOC in thiswatershed. 13C values of soil atmosphereCO2 (–17.2 to –25.2) were slightly lessnegative than would be expected for production viaaerobic soil microbial decomposition and rootrespiration. There was a close correspondence between13C values (–15.5 to –21.5) forstreamwater TIC and soil atmospheric CO2 in thecentral part of the catchment where the stream drainsCO2-rich peats. 14C dating showed thatalthough peat has been accumulating in the watershedfor at least 2700 years, DOC in soil pore water andstreamwater contains carbon of predominantly recentorigin (post-AD 1955).  相似文献   

12.
Conversion of methanol to CH4 has a large isotope effect so that a small contribution of methanol-dependent CH4 production may decrease the 13CH4 of total CH4 production. Therefore, we investigated the role of methanol for CH4 production. Methanol was not detectable above 10 M in anoxic methanogenic rice field soil. Nevertheless, addition of 13C-labeled methanol (99% enriched) resulted in immediate accumulation of 13CH4. Addition of 0.1 M 13C-methanol resulted in increase of the 13CH4 from –47 to –6 within 2 h, followed by a slow decrease. Addition of 1 M 13C-methanol increased 13CH4 to +500 within 4 h, whereas 10 M increased 13CH4 to +2500 and continued to increase. These results indicate that the methanol concentrations in situ, which diluted the 13C-methanol added, were 0.1 M and that the turnover of methanol contributed only about 2% to total CH4 production at 0.1 M. However, contribution increased up to 5 and 17% when 1 and 10 M methanol were added, respectively. Anoxic rice soil that was incubated at different temperatures between 10 and 37 °C exhibited maximally 2–6% methanol-dependent methanogenesis about 1–2 h after addition of 1 M 13C-methanol. Only at 50 °C, contribution of methanol to CH4 production reached a maximum of 10%. After longer (7–10 h) incubation, however, contribution generally was only 2–4%. Methanol accumulated in the soil when CH4 production was inhibited by chloroform. However, the accumulated methanol accounted for only up to 0.7 and 1.2% of total CH4 production at 37 and 50 °C, respectively. Collectively, our results show that methanol-dependent methanogenesis was operating in anoxic rice field soil but contributed only marginally to total CH4 production and the isotope effect observed at both low and high temperature.  相似文献   

13.
Opioid agonists specific for the , , and opioid receptor subtypes were tested for their ability to modulate potassium-evoked release of L-glutamate and dynorphin B-like immunoreactivity from guinea pig hippocampal mossy fiber synaptosomes. The opioid agonists U-62,066E and (–) ethylketocyclazocine, but not the agonist [D-Ala2,N-MePhe4,Gly5-ol]-enkephalin (DAGO) nor the agonist [D-Pen2,5]enkephalin (DPDE), inhibited the potassium-evoked release of L-glutamate and dynorphin B-like immunoreactivity. U-62,066E, but not DAGO or DPDE, also inhibited the potassium-evoked rise in mossy fiber synaptosomal cytosolic Ca2+ levels, indicating a possible mechanism for agonist inhibition of transmitter release. DAGO and DPDE were found to be without any effect on cytosolic Ca2+ levels or transmitter release in this preparation. The U-62,066E inhibition of the potassium-evoked rise in synaptosomal cytosolic Ca2+ levels was partially attenuated by the opioid antagonist quadazocine and insensitive to the -opioid specific antagonist ICI 174,864 and the opioid-preferring antagonists naloxone and naltrexone. Quadazocine also reversed U-62,066E inhibition of the potassium-evoked release of L-glutamate, but not dynorphin B-like immunoreactivity. These results suggest that opioid agonists inhibit transmitter release from mossy fiber terminals through both opioid and non- opioid receptor mediated mechanisms.  相似文献   

14.
Summary Foliar samples were obtained from symbiotic nitrogen-fixers and control plants (non-fixers) along elevational and primary successional gradients in volcanic sites in Hawai'i. Most control plants had negative 15N values (range-10.1 to +0.7), while most nitrogen-fixers were near 0. Foliar 15N in the native tree Metrosideros polymorpha did not vary with elevation (from sea level to tree-line), but it did increase substantially towards 0 on older soils. The soil in an 197-yr-old site had a 15N value of approximately-2, while in a 67000-yr-old site it was +3.6. We suggest that inputs of 15N-depleted nitrogen from precipitation coupled with very low nitrogen outputs cause the strongly negative 15N values in non-nitrogen-fixing plants on early successional sites.  相似文献   

15.
Three species of the reef coral genus Madracis display skeletal isotopic characteristics that relate to depth, colony topography, and consequently to coral physiology. The joint interpretation of skeletal 13C and 18O provides information on the ecological plasticity and adaptation to depth of a coral species. Isotopic results are most easily understood in terms of kinetic effects, which reduce both 18O and 13C below isotopic equilibrium values, and metabolic effects, which only influence the skeletal 13C. Madracis mirabilis is adapted to depths shallower than 20 m, and shows the greatest range in kinetic effects and the strongest metabolic 13C enrichments caused by symbiont photosynthesis. Madracis formosa lives deeper than 40 m, and shows a reduced range of kinetic effects and relatively weak metabolic 13C enrichments. Madracis pharensis inhabits depths from 5 to >60 m, and does not attain the strength of kinetic effects of either of the other two species, apparently because it is not quite as well adapted to rapid growth at either extreme.  相似文献   

16.
Perchloric acid extracts were prepared from liquid-N2-frozen gerbil and guinea pig brain slices studied under one of three conditions: O2-incubated, N2-incubated or O2-incubated recovery following N2 incubation. Mole percentages of the various phosphatic components contained in the extracts were determined by phosphorus-31 nuclear magnetic resonance spectroscopy. The brain slice extract spectrum revealed a previously unreported group of brain phosphodiesters at –0.73 relative to 85% orthophosphoric acid Although the phosphatic profiles from O2-incubated slices fromgerbils and guinea pigs revealed only minor species variations, which differed quantitatively rather than qualitatively, species-specific differences were made readily apparent and amplified by incubating brain slices under oxygen-deficient conditions. Despite these differences which were most prevalent during the recovery phase, the overall metabolic changes described herein in response to N2-incubation were in accord with the results obtained by other analytical techniques. Inorganic orthophosphate (2.63 ) was increased, while nucleoside (principally, adenosine) triphosphate (-, –10.92 , -, –21.45 , and -, –5.80 ) and phosphocreatine (–3.12 ) levels were decreased in response to N2 incubation. In addition, inosine monophosphate (3.78 ) was increased and the levels of a partially characterized acid-labile phosphate (0.85 , guinea pig) were decreased upon N2 incubation. Phosphoglyceride metabolism also appeared to be altered by oxygen deprivation (gerbil). These latter findings provide additional information concerning the metabolic responses of cerebral tissue to oxygen deficient conditions.Dedicated to Henry McIlwain.  相似文献   

17.
Natural abundance of 15N in tropical plants with emphasis on tree legumes   总被引:6,自引:0,他引:6  
Natural abundance of 15N ( 15N) of leaves harvested from tropical plants in Brazil and Thailand was analyzed. The 15N values of non-N2-fixing trees in Brazil were +4.5±1.9, which is lower than those of soil nitrogen (+8.0±2.2). In contrast, mimosa and kudzu had very low 15N values (–1.4+0.5). The 15N values of Panicum maximum and leguminous trees, except Leucaena leucocephala, were similar to those of non-N2-fixing trees, suggesting that the contribution of fixed N in these plants is negligible. The 15N values of non-N2-fixing trees in Thailand were +4.9±2.0. Leucaena leucocephala, Sesbania grandiflora, Casuarina spp. and Cycas spp. had low 15N values, close to the value of atmospheric N2 (0), pointing to a major contribution of N2 fixation in these plants. Cassia spp. and Tamarindus indica had high 15N values, which confirms that these species are non-nodulating legumes. The 15N values of Acacia spp. and Gliricidia sepium and other potentially nodulating tree legumes were, on average, slightly lower than those of non-N2-fixing trees, indicating a small contribution of N2 fixation in these legumes.  相似文献   

18.
Even though the genomes of several major species have been sequenced, many orphan receptors with unknown ligands and mechanisms of action remain in the CNS. The 2 glutamate receptor (GluR2) is one of such receptors expressed predominantly in the cerebellar Purkinje cells. On the basis of amino acid similarity, it belongs to ionotropic glutamate receptor (iGluR) family, which mediates fast excitatory neurotransmission in the mammalian CNS. Although its null-mutant mice show prominent motor discoordination, the mechanisms by which GluR2 participates in the cerebellar functions have been unclear. To gain insight into GluR2s mechanisms, we recently generated mice that express either a wild-type or a mutant GluR2 transgene, in which the conserved arginine in GluR2s N-terminal putative ligand-binding motif was disrupted. By breeding these transgenic mice onto a GluR2–/– background, we obtained two transgenic rescue lines. Surprisingly, the mutant GluR2 transgene was as effective as the wild-type GluR2 in rescuing the GluR2-null mice. As the disrupted arginine residue is highly conserved from ancestral bacterial periplasmic amino acid-binding proteins to mammalian iGluRs, we propose that GluR2 may not require glutamate-like amino acids and may function in an unconventional manner. This transgenic rescue approach to investigating orphan receptors is a relatively easy but powerful method when a knockout mouse with a distinct phenotype is already available. The advantages and limitations of this approach, together with certain cautions in interpreting the resulting data, are discussed in this review.  相似文献   

19.
In a series of experiments, it is demonstrated that the trace element vanadium (4·10-7 g-at/l as NH4VO3) has a considerable positive influence on the synthesis of -aminolevulinic acid (-ALA) in the autotrophically growing green algaChlorella pyrenoidosa, the effect being visible by an enhanced output of the amino acid into the culture medium in presence of levulinic acid (LA). The level of intracellularly accumulated -ALA, however, is not changed in presence of the metal. The V-effect on exogenous found -ALA is suppressed, when LA is added to the nutrient medium at low pH (pH 5), although V-uptake into the algal cells is not disturbed by LA. As demonstrated in culture media with various nitrogen sources (urea, partially hydrolized urea, ammonium salts), the development of the pH during the cultivation time is important for the presentation of the V-effect on -ALA. It is suggested that vanadium acts as a catalyst in the conversion of 4,5-dioxovaleric acid to -ALA by transamination.Abbreviations -ALA -aminolevulinic acid - LA levulinic acid - DOVA 4,5-dioxovaleric acid  相似文献   

20.
Given a uniform N source, the 15N of barley shoots provided a genotypic range within treatments and a separation between control and salt-stress treatments as great as did 13C*. Plant 15N has been represented in the literature as a bioassay of external source 15N and used to infer soil N sources, thus precluding consideration of the plant as a major cause in determining its own 815N. We believe this to be the first report of plant 15N as a genetic trait. No mechanistic model is needed for use of 15N as a trait in controlled studies; however, a qualitative model is suggested for further testing.Symbol 15N (or 13C) the difference between: (1) the ratio of heavy to light isotopes of the element in a sample and (2) that of its reference standard  相似文献   

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