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21.
Glutamine and Aspartate Loading of Synaptosomes: A Reevaluation of Effects on Calcium-Dependent Excitatory Amino Acid Release 总被引:5,自引:3,他引:2
Guinea-pig cerebral cortical synaptosomes were preincubated for 60 min with 100 microM D-aspartate, L-aspartate, or L-glutamate. The total D- plus L-aspartate content of the synaptosomal fraction increased to 235%, 195%, or 164%, respectively, of the control. Despite this no increase was seen in the very low KCl evoked, Ca2+-dependent release of aspartate. Preincubation with the three amino acids changed the synaptosomal glutamate content to 78% (D-aspartate), 149% (L-aspartate), or 168% (L-glutamate) of control. However there was no statistically significant effect of these preincubations on the extent of Ca2+-dependent glutamate release. Thus the Ca2+-dependent release of aspartate and glutamate is not determined by the total synaptosomal content of these amino acids. The addition of 0.1-0.5 mM glutamine to the incubation caused a massive appearance of glutamate in the extrasynaptosomal medium. Analysis of specific activities showed that glutamine was hydrolysed directly by an extrasynaptosomal glutaminase, and that intrasynaptosomal glutamate was predominantly labelled by uptake of this glutaminase-derived glutamate. No increase was seen in the extent of Ca2+-dependent release of glutamate (by fluorimetry) either after preincubation with glutamine or in the continued presence of glutamine. Thus we are unable to confirm reports that glutamine expands the transmitter pool of glutamate. The extrasynaptosomal glutaminase activity in the synaptosomal preparation was inhibited by Ca2+ and activated by phosphate. Identical kinetics were obtained with "free" brain mitochondria, confirming the origin of the glutamine-derived glutamate. 相似文献
22.
Replicon rescue: a novel strategy to clone the genomic DNA flanking insertions of integrating shuttle vector DNA.
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T L McMahon Z Wilczynska C Barth D J Fraser L Pontes P R Fisher 《Nucleic acids research》1996,24(20):4096-4097
A novel cloning strategy, replicon rescue, was developed for cloning genes disrupted by plasmid insertions. After ligation to a tetracycline resistance cassette, fragments containing a bacterial origin of replication from the insertion are recovered in Escherichia coli because they replicate autonomously. Restriction enzymes for cloning are so chosen that the only legitimate two fragment ligation yielding TetR clones involves a fragment spanning the boundary of the insertion. Replicon rescue was used successfully firstly in a test system to clone the chromosomal orl from a Klebsiella aerogenes strain, and secondly to recover a disrupted gene from a phototaxis-deficient mutant of Dictyostelium. 相似文献
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Daniel Zuleta Gabriel Arellano Sean M. McMahon Salomón Aguilar Sarayudh Bunyavejchewin Nicolas Castaño Chia-Hao Chang-Yang Alvaro Duque David Mitre Musalmah Nasardin Rolando Pérez I-Fang Sun Tze Leong Yao Renato Valencia Sruthi M. Krishna Moorthy Hans Verbeeck Stuart J. Davies 《Global Change Biology》2023,29(12):3409-3420
Accurate estimates of forest biomass stocks and fluxes are needed to quantify global carbon budgets and assess the response of forests to climate change. However, most forest inventories consider tree mortality as the only aboveground biomass (AGB) loss without accounting for losses via damage to living trees: branchfall, trunk breakage, and wood decay. Here, we use ~151,000 annual records of tree survival and structural completeness to compare AGB loss via damage to living trees to total AGB loss (mortality + damage) in seven tropical forests widely distributed across environmental conditions. We find that 42% (3.62 Mg ha−1 year−1; 95% confidence interval [CI] 2.36–5.25) of total AGB loss (8.72 Mg ha−1 year−1; CI 5.57–12.86) is due to damage to living trees. Total AGB loss was highly variable among forests, but these differences were mainly caused by site variability in damage-related AGB losses rather than by mortality-related AGB losses. We show that conventional forest inventories overestimate stand-level AGB stocks by 4% (1%–17% range across forests) because assume structurally complete trees, underestimate total AGB loss by 29% (6%–57% range across forests) due to overlooked damage-related AGB losses, and overestimate AGB loss via mortality by 22% (7%–80% range across forests) because of the assumption that trees are undamaged before dying. Our results indicate that forest carbon fluxes are higher than previously thought. Damage on living trees is an underappreciated component of the forest carbon cycle that is likely to become even more important as the frequency and severity of forest disturbances increase. 相似文献
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Two intertidal snails, Littorina saxatilis (Olivi, 1972) (upper eulittoral fringe/maritime zone) and Littorina obtusata (Linnaeus, 1758) (lower eulittoral) were collected from a boulder shore on Nobska Point, Cape Cod, Massachusetts, in July
and acclimated for 15–20 days at 4 ° or 21 °C. Oxygen consumption rate (Vo2) was determined for 11–15 subsamples of individuals at 4 °, 11 ° and 21 °C with silver/platinum oxygen electrodes. Multiple
factor analysis of variance (MFANOVA) of lo10 transformed values of whole animal Vo2 with log10 dry tissue weight (DTW) as a covariant revealed that increased test temperature induced a significant increase in Vo2 in both species (P<0.00001). In contrast, MFANOVA revealed that temperature acclimation did not affect Vo2 in either L. saxatilis (P= 0.35) or L. obtusata (P= 0.095). Thus, neither species displayed a capacity for the typical metabolic temperature compensation marked by an increase
in Vo2 at any one test temperature in individuals acclimated to a lower temperature that is characteristic of most ectothermic animals.
Lack of capacity for metabolic temperature acclimation has also been reported in other littorinid snail species, and may be
characteristic of the group as a whole. Lack of capacity for respiratory temperature acclimation in these two species and
other littorinids may reflect the extensive semi-diurnal temperature variation that they are exposed to in their eulittoral
and eulittoral fringe/maritime zone habitats. In these habitats, any metabolic benefits derived from longer-term temperature
compensation of metabolic rates are negated by extreme daily temperature fluctuations. Instead, littorinid species appear
to have evolved mechanisms for immediate metabolic regulation which, in L. saxatilis and L. obtusata and other littorinids, appear to centre on a unique ability for near instantaneous suppression of metabolic rate and entrance
into short-term metabolic diapause at temperatures above 20–35 °C, making typical seasonal respiratory compensation mechanisms
characteristic of most ectotherms of little adaptive value to littorinid species. 相似文献
27.
Nitrogenous excretion in two snails, Littorina saxatilis (high intertidal) and L. obtusata (low intertidal) was studied in relation to temperature acclimation (at 4° and 21°C), including total N excretion rates,
the fraction of urea in N excretion, corresponding O:N ratios and the partitioning of deaminated protein between catabolic
and anabolic processes at 4°, 11° and 21°C. Aggregate N excretion rates in both species showed no significant compensatory
adjustments following acclimation. Total weight specific N excretion rates at 21°C were higher in standard 3 mg L. saxatilis (739 ng N mg−1 h−1) than standard 5 mg L. obtusata (257 ng N mg−1 h−1) for snails acclimated to 21°C. Comparisons of Q10 values of total weight specific N excretion to Q10 values for weight specific oxygen consumption ({xxV}O2) between 4° to 11 °C and 11° to 21°C indicated that, while total rates of catabolic metabolism ({xxV}O2) and protein deamination in L. obtusata were essentially parallel, the relationship between N excretion and {xxV}O2 in L. saxatilis revealed the partitioning of a larger share of deaminated protein carbon into anabolism at 4° and 21°C than at 11°C. Urea
N accounted for a larger share of aggregate N excreted in L. saxatilis than in L. obtusata, but in both species urea N is a greater proportion of total N excreted when acclimated at 4°C (urea N: ammonia N ratio range:
1 to 2.15) than in snails acclimated to 21°C (urea N: ammonia N ratio range: 0.46 to 1.39). Molar O:N ratios indicate that
the proportion of metabolism supported by protein catabolism is greater in L. saxatilis (O:N range: 2.5–8.4) than in L. obtusata (O:N range: 7.3–13.0). In both species, regardless of acclimation temperature, the O:N ratios are generally lowest (high
protein catabolism) at 4°C and highest at 21°C. 相似文献
28.
McMahon Jennifer M.; White Wanda L.B.; Sayre Richard T. 《Journal of experimental botany》1995,46(7):731-741
Cassava is the most agronomically important of the cyanogeniccrops. Linamarin, the predominant cyanogenic glycoside in cassava,can accumulate to concentrations as high as 500 mg kg1fresh weight in roots and to higher levels in leaves. Recently,the pathway of linamarin synthesis and the cellular site oflinamarin storage have been determined. In addition, the cyanogenicenzymes, linamarase and hydroxynitrile lyase, have been characterizedand their genes cloned. These results, as well as studies onthe organ- and tissue-specific localization of linamarase andhydroxy-nitrile lyase, allow us to propose models for the regulationof cyanogenesis in cassava. There remain, however, many unansweredquestions regarding the tissue-specific synthesis, transport,and accumulation of cyanogenic glycosides. The resolution ofthe sequestions will facilitate the development of food processing,biochemical and transgenic plant approaches to reducing thecyanogen content of cassava foods. Key words: Cyanide, cyanogenic glycosides, linamarin, cyanogens 相似文献
29.
Foraging success of largemouth bass at different light intensities: implications for time and depth of feeding 总被引:2,自引:0,他引:2
Laboratory feeding trials were conducted to determine how light intensity affects foraging success by the visual piscivore, the largemouth bass ( Micropterus salmoides ). Foraging success was greater than 95% at light levels ranging from low intensity daylight (2.43 × 102 lx) to moonlight (3 × 10−3 lx), but declined significantly to 62% at starlight (2 × 10−4 lx) and near 0% in total darkness. Over a range of low to high water clarities (0.5, 2.0, and 4.0 m Secchi depth), estimated depth limits for feeding during the day ranged between 5.5 to 44 m and from 1.6 to 13 m at night during a full moon. At starlight, light intensity rapidly attenuated to a level below the feeding threshold within 0.5 m of the surface at all water clarities. The depth of the water column available for feeding in low clarity water (0.5 m Secchi) was 67 and 75% less than at moderate (2.0 m Secchi) and high (4.0 m Secchi) water clarities. The findings illustrate how differences in the light environment can have important ramifications for predator-prey interactions. 相似文献
30.
The influence of track compliance on running 总被引:5,自引:0,他引:5
A model of running is proposed in which the leg is represented as a rack-and-pinion element in series with a damped spring. The rack-and-pinion element emphasizes the role of descending commands, while the damped spring represents the dynamic properties of muscles and the position and the rate sensitivity of reflexes. This model is used to predict separately the effect of track compliance on step length and ground contact time. The predictions are compared with experiments in which athletes ran over tracks of controlled spring stiffness. A sharp spike in foot force up to 5 times body weight was found on hard surfaces, but this spike disappeared as the athletes ran on soft experimental tracks. Both ground contact time and step length increased on very compliant surfaces, leading to moderately reduced running speeds, but a range of track stiffness was discovered which actually enhances speed. 相似文献