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891.
J. V. Priestley M. A. Hynes V. K. M. Han M. Réthelyi E. R. Perl P. K. Lund 《Histochemistry and cell biology》1988,89(5):467-479
Summary Methodological variables for in situ hybridization using 32P labelled oligodeoxyribonucleotides (oligomers) have been examined. Four different oligomers directed against proglucagon messenger RNA (mRNA) and two different oligomers against prosomatostatin mRNA have been used. Specific hybridization was obtained in adult rat brain, stomach and pancreas and in neonatal rat ileum. Tissue was perfusion fixed with 4% paraformaldehyde 0.2% glutaraldehyde and hybridization was carried out in 50% formamide for 72 h at 42° C. Using hybridization conditions of lower stringency (33% formamide) labelling was also obtained in guinea pig tissue. Other variables which affected hybridization signal intensity were the inclusion of a prehybridization dehydration stage, the probe concentration, the inclusion of ammonium acetate in the posthybridization dehydrating ethanols and in the autoradiographic emulsion, and the exposure time. The localisation of proglucagon mRNA in rat pancreas using a 20mer was used as a model tissue for testing these methodological variables and the results were found generally also to apply to the other probes and tissues tested. The methods described provide single cell resolution and show that 32P labelled oligomers may be used to localise neuropeptide and endocrine mRNAs in different types of tissue and in different mammalian species. 相似文献
892.
Summary In order to study mitogenic control during axolotl limb regeneration, we have developed a primary blastema cell culture as a very sensitive bioassay for blastema mitogens. Transferrin, an iron-binding glycoprotein which has been shown to be the neurotrophic factor for muscle cells, is the mitogen which has been analysed in the present report. Addition of approximately 2 g human transferrin/ ml of serum-free culture medium enhances blastema cell proliferation 11-fold over control levels and 2-fold over that produced by the addition of nerve extracts or purified growth factors extracted from nerve tissues (basic and acidic fetal growth factor, FGF). At a higher concentration (20 g/ml), transferrin alone has no mitogenic effect unless the medium is also supplemented with FeCl3 (100 M). The results are discussed with regard to the sensitivity of the blastema cell culture bioassay and in the context of the neurotrophic theory of urodele limb regeneration. 相似文献
893.
Summary Phenotypic analyses of genetic combinations involving the gene extramacrochaetae (emc) reveal its participation in the differentiation of both sensory elements and wing veins. The study of near-amorphic alleles of emc in mitotitc recombination clones indicates that it also affects cell proliferation. These clones show abnormal sizes, shapes and spatial distribution. They differentiate extra sensory elements as well as extra veins. A gain of function mutation in the gene causes opposite phenotypes in both differentiation systems. The effects of the mutant on proliferation and patterning are consistent with the emc gene being involved in the transfer of information between neighbouring cells, which leads to the spatial expression of the achaetescute gene complex and genes involved in vein formation. 相似文献
894.
J. Šťovíčková J. Turková B. Urbánek V. Bažant J. Kavalírová 《Biotechnology Techniques》1988,2(2):121-126
Summary The effect was studied of the method of drying chymotrypsin attached to bead cellulose on its chemical and physical characteristics. These characteristics are not deteriorated when replacing the commonly used lyophilisation by fluid drying in the air stream. The preparations saved essentially the same proteolytic activity as well as stability even when increasing the drying air temperature to 70°C. 相似文献
895.
Summary By means of an experimental factorial design the role of several culture conditions is described and its values determined for the maximum yeast production in mixed cultures ofAspergillus oryzae andRhodotorula glutinis. 相似文献
896.
897.
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
898.
899.
M. Pilar López M. Jose Gómez-Lechón Jose V. Castell 《In vitro cellular & developmental biology. Plant》1988,24(6):511-517
Summary This study examines the factors involved in the rapid glycolysis and glycogenolysis that occur during the first stages of
hepatocyte culture: a) Shortly after seeding glycolysis, estimated as lactate released to culture medium, increased 10 times
in comparison to that reported in vivo. By 8 to 9 h of culture, hepatocytes were nearly glycogen-depleted even in the presence
of insulin. b) 6-Phosphofructo-2-kinase remained 100% active during this period. The proportion of the initial active phosphorylase
(87%) decreased to 57% by 7 h of culture. c) Fructose 2,6-bisphosphate content was initially similar to that found in liver
of fed animals, decreased after seeding and increased thereafter up to four times the initial concentration. In spite of changes
in the concentration of this activator, the glycolytic rate remained high and constant. d) ADP and AMP increased sharply after
cell plating, reaching values 1.7 and 3.5 times higher. The rise in AMP levels may be involved in the activation of glycolysis
and glycogenolysis, because this metabolite is known to act as an allosteric activator of phosphofrucktokinase and glycogen
phosphorylase. This metabolic situation resembles that of cells under hypoxia.
Part of this work was presented at the 38th Annual Meeting of the Tissue Culture Association, Washington, DC, May 1987. 相似文献
900.
In order to improve the interpretation of the results of pollen analyses concerning mediaeval anthropogenous sediments a set of recent seeds of the principal cereals was pollenanalytically elaborated. The aim was to find out whether pollen grains can be preserved on mature seeds. The pollen spectra obtained from the material of mediaeval anthropogenous sediments include, in most cases, a large amount of pollen of certain plants including cereals. It is suggested that part of the pollen spectra from anthropogenous sediments need not come from air transport, i.e. from the surrounding vegetation; it could get into waste pits in another way. Above all, some foodstuffs were considered to be the sources of pollen grains. The origin of some grains coming from honey and from plant infusions has, at least partly, been demonstrated (Jankovská 1987). As the next step, attention was paid to other products, mainly those of cereals. An experimental pollen analysis of recent seeds of selected cereals showed that pollen grains are preserved on their surface, often in high concentrations. This is due, above all, to the cleistogamy of most cereals. The highest values of pollen grains have so far been found inAvena sativa andPanicum milliaceum. Triticum aestivum andHordeum vulgare showed lower values of pollen grains. The lowest amounts were found in heterogamousSecale cereale. Simultaneously, the pollen of plants growing in a neighbourhood, above all the pollen of field weeds, was found sticking to the seeds. 相似文献