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51.
Development and characterization of continuous avian cell lines depleted of mitochondrial DNA 总被引:6,自引:0,他引:6
Réjean Morais Paul Desjardins Chanta Turmel Karen Zinkewich-Péotti 《In vitro cellular & developmental biology. Plant》1988,24(7):649-658
Summary Populations of quail and chicken cells were treated with ethidium bromide, an inhibitor of mitochondrial DNA replication.
After long-term exposure to the drug, the cell populations were transferred to ethidium bromide (EtdBr)-free medium, and cloned.
Clones HCF7 (quail) and DUS-3 (chicken) were propagated for more than a year, and then characterized. Analysis of total cellular
DNA extracted from these cells revealed no characteristic mitochondrial DNA molecule by Southern blot hybridization of HindIII-
or AvaI-digested total cellular DNA probed with cloned mitochondrial DNA fragments. Reconstruction experiments, where a small
number of parental cells was mixed with HCF7 cells and DUS-3 cells before extraction of total cellular DNA, further strengthen
the notion that the drug-treated cells are devoid of mitochondrial DNA molecules. The cell populations were found to proliferate
at a moderately reduced growth rate as compared to their respective parents, to be auxotrophic for uridine, and to be stably
resistant to the growth inhibitory effect of EtdBr and chloramphenicol. At the ultrastructural level, mitochondria were considerably
enlarged and there was a severe reduction in the number of cristae within the organelles and loss of cristae orientation.
Morphometric analysis revealed a fourfold increase of the mitochondrial profile area along with a twofold decrease of the
numerical mitochondrial profiles. Analysis of biochemical parameters indicated that the cells grew with mitochondria devoid
of a functional respiratory chain. The activity of the mitochondrial enzyme dihydroorotate dehydrogenase was decreased by
95% and presumably accounted for uridine auxotrophy.
This work was supported by a grant from the Medical Research Council of Canada. 相似文献
52.
53.
Charles R. Ill Tammy Brehm Bjorn K. Lydersen Rachel Hernandez Karen G. Burnett 《In vitro cellular & developmental biology. Plant》1988,24(5):413-419
Summary Studies with Human x Human (HxH), Human x Mouse (HxM), and Mouse x Mouse (MxM) hybridomas have enabled us to define specific
factors that affect hybridoma growth in a species-specific manner. Three transferrins and three lipophilic iron chelates have
been tested for their ability to support hybridoma proliferation and antibody production. The results of these studies demonstrate
that HxH hybridomas do not respond to bovine transferrin a+ concentrations up to 100 μg/ml and are approximately 100-fold less responsive to mouse transferrin than to human transferrin.
HxM and MxM hybridomas respond equally to human or mouse transferrin but are 100-fold less sensitive to bovine transferrin.
An antibody to the human transferrin receptor inhibited the growth-promoting activity of human or mouse transferrin on HxH
hybridomas but was ineffective on HxM hybridomas. This semonstrated the functionality of the human transferrin receptor in
HxH hybridomas and that human, mouse, and bovine transferrin were interacting through the mouse transferrin receptor in HxM
hybridomas. HxH and HxM hybridomas respond similarly to three different iron chelates exhibiting 80 to 110% of the growth
response to human transferrin. MxM hybridomas fail to respond to the iron chelates at similar concentrations, suggesting that
the human genome present in the other hybridoma species confers a unique ability for utilizing iron when delivered in this
form. 相似文献
54.
Sasha N. Zill S. Faith Frazier John Lankenau Karen Jepson-Innes 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1992,170(6):761-772
Summary The use of the locust (Schistocerca americana) hindleg in postural control was examined in animals that stood on a repeatedly swayed vertical substrate. Myograms were recorded from leg muscles and the angle of the femoro-tibial joint was monitored photographically. Two discrete strategies were observed,; in compensatory reactions the hindleg was held in place, while in flexion reactions, the leg was moved, most often to complete flexion of the femoro-tibial joint. Tightly coupled, rhythmic bursting occurred in the flexor and levator muscles of the leg during compensatory reactions. Bursting was initiated repeatedly when the substrate was being pulled away from the animal. Bursting was correlated with subsequent decreases in the rate of change of the femorotibial joint angle. Compensatory and flexion reactions occurred preferentially in different ranges of joint angles: most often, compensatory reactions occurred in the midrange, while flexion reactions were elicited in the extremes of joint angle. These differences may be due to the mechanical advantages of the tibial muscles and the leg may be moved to full flexion because of a locking mechanism of the flexor muscle tendon. These reactions are compared with known reflexes of hindleg proprioceptors and contrasted with similar responses of vertebrates. 相似文献
55.
Hypoxanthine-guanine phosphoribosyltransferase (HPRT, EC 2.4.2.8) is a purine salvage enzyme that catalyses the conversion of hypoxanthine and guanine to their respective mononucleotides. Partial deficiency of this enzyme can result in the overproduction of uric acid leading to a severe form of gout, whilst a virtual absence of HPRT activity causes the Lesch-Nyhan syndrome which is characterised by hyperuricaemia, mental retardation, choreoathetosis and compulsive self-mutilation. The HPRT-encoding gene is located on the X chromosome in the region q26–q27 and consists of nine exons and eight introns totalling 57 kb. This gene is transcribed to produce an mRNA of 1.6 kb, which contains a protein encoding region of 654 nucleotides. With the advent of increasingly refined techniques of molecular biology, it has been possible to study the HPRT gene of individuals with a deficiency in HPRT activity to determine the genetic basis of the enzyme deficiency. Many different mutations throughout the coding region have been described, but in the absence of precise information on the three-dimensional structure of the HPRT protein, it remains difficult to determine any consistent correlation between the structure and function of the enzyme. 相似文献
56.
57.
The regulation of PAG by ammonia in whole brain (Sprague-Dawley) and regional (Fischer-344) synaptosomal preparations from adult and aged animals was assessed. Whole brain synaptosomal preparations from both age groups displayed a significant decrease in PAG activity with increasing ammonium chloride concentrations, however, the aged rats exhibited a significant attenuation in ammonia-induced PAG inhibition. PAG activity measured in synaptosomes prepared from the striatum (STR), temporal cortex (TCX) and hippocampus (HIPP) was also inhibited by ammonium chloride. The STR showed the greatest degree of ammonia-induced PAG inhibition (55%) followed by the HIPP (30–35%) and the TCX (25–30%). This reduction in PAG activity was significantly attenuated in STR from aged rats at ammonium chloride concentrations greater than 50 M and in the TCX, PAG activity was significantly attenuated in the aged rats at ammonia concentrations of 0.5 and 1.0 mM. Ammonia regulation of PAG activity in the HIPP appeared to be unaffected by age. Ammonium chloride concentrations up to 5 mM had no effect on GLU release from cortical slices, although GLN efflux was significantly enhanced. These findings suggest that isozymes of PAG may exist in different brain regions based on their differential sensitivity to ammonia. The attenuation of ammonia-induced PAG inhibition seen in aged rats may have deleterious effects in the aged brain.Abbreviations PAG
phosphate-activated glutaminase: L-glutamine amidohydrolase; EC 3.5.1.2
- STR
striatum
- TCX
temporal cortex
- HIPP
hippocampus 相似文献
58.
Seasonal carbon isotope discrimination in a grassland community 总被引:11,自引:0,他引:11
Mark P. Smedley Todd E. Dawson Jonathan P. Comstock Lisa A. Donovan Dorothy E. Sherrill Craig S. Cook James R. Ehleringer 《Oecologia》1991,85(3):314-320
Summary Grassland communities of arid western North America are often characterized by a seasonal increase in ambient temperature and evaporative demand and a corresponding decline in soil moisture availability. As the environment changes, particular species could respond differently, which should be reflected in a number of physiological processes. Carbon isotope discrimination varies during photosynthetic activity as a function of both stomatal aperture and the biochemistry of the fixation process, and provides an integrated measure of plant response to seasonal changes in the environment. We measured the seasonal course of carbon isotope discrimination in 42 grassland species to evaluate changes in gas exchange processes in response to these varying environmental factors. The seasonal courses were then used to identify community-wide patterns associated with life form, with phenology and with differences between grasses and forbs. Significant differences were detected in the following comparisons: (1) Carbon isotope discrimination decreased throughout the growing season; (2) perennial species discriminated less than annual species; (3) grasses discriminated less than forbs; and (4) early flowering species discriminated more than the later flowering ones. These comparisons suggested that (1) species active only during the initial, less stressful months of the growing season used water less efficiently, and (2) that physiological responses increasing the ratio of carbon fixed to water lost were common in these grassland species, and were correlated with the increase in evaporative demand and the decrease in soil moisture. 相似文献
59.
Summary We investigated the effects of thorns and spines on the feeding of 5 herbivore species in arid Australia. The herbivores were the rabbit (Oryctolagus cuniculus), euro kangaroo (Macropus robustus), red kangaroo (Macropus rufus), sheep (Ovis aries), and cattle (Bos taurus). Five woody plants without spines or thorns and 6 woody plants with thorns were included in the study. The spines and thorns were not found to affect the herbivores' rates of feeding (items ingested/min), but they did reduce the herbivores' rates of biomass ingestion (g-dry/item). The reduction in biomass ingested occurred in two ways: at a given diameter, twigs with spines and thorns had less mass than undefended plants, and the herbivores consumed twigs with smaller diameters on plants with spines and thorns. The relative importance of the two ways that twigs with spines and thorns provided less biomass varied with herbivore body mass. Reduced twig mass was more important for small herbivores, while large herbivores selected smaller diameters. The effectiveness of spines and thorns as anti-herbivore defenses did not vary with the evolutionary history of the herbivores (i.e. native vs. introduced). Spines and thorns mainly affected the herbivores' selection of maximum twig sizes (reducing diameter and mass), but the minimum twig sizes selected were also reduced. 相似文献
60.
Grande Karen D.; Bender Michael L.; Irwin Brian; Platt Trevor 《Journal of plankton research》1991,13(1):1-16
In vitrorates of gross and net oxygen production were measuredas a function of light intensity in some plankton communitiescollected from Bedford Basin, Nova Scotia, and in a monoclonalculture of Synechococcus. The rate of gross oxygen productionwas measured by a technique in which the stable oxygen isotope,18O, serves as a photosynthetic tracer Net oxygen productionwas measured by automated Winkler technique. The rate of communityrespiration in the light was then determined by the differencebetween gross and net rates of oxygen production. In the naturalpopulations examined, neither gross nor net oxygen productionrates were significantly inhibited at the highest light intensitymeasured (500800 µE m2 s1) In a samplein which the dark respiration rate was small relative to themaximal rate of production [Pmax;sensu Platt et al (1980) JMar. Res., 38, 687701] the rates of lightrespiration were 3 times greater. In two other communities,with high rates of dark respiration relative to Pmaxthe ratesof light respiration were closer to rates of darkrespiration. In the Synechococcus clone, both gross and netoxygen production rates were inhibited at high light intensities.Rates of light respiration were found to varyas a function of light intensity. The greatest rates of respirationwere measured in samples incubated at light intensities thatwere just saturating (100 µE m2 s1). Therates of 14C production were also measured as a function oflight intensity The photosynthetic quotients, based on 14C productionrates and gross oxygen production rates, average 1 9 相似文献