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61.
Abstract: The relationship between elevations in intracellular free Ca2+ concentration ([Ca2+]i) by different mechanisms and tyrosine hydroxylase (TH) gene expression was examined. Depolarization by an elevated K+ concentration triggered rapid and sustained increases in [Ca2+]i from a basal level of ~50 to 110–150 nM and three- to fourfold elevations in TH mRNA levels, requiring extracellular calcium but not inositol 1,4,5-trisphosphate (IP3). On the other hand, bradykinin or thapsigargin, both of which induce release of intracellular calcium stores via IP3 or inhibition of Ca2+-ATPase, rapidly elevated [Ca2+]i to >200 nM and increased TH gene expression (three-to fivefold). Confocal imaging showed that the elevations in [Ca2+]i in each case occurred throughout the cyto- and nucleoplasm. The initial rise in [Ca2+]i due to either bradykinin or thapsigargin, which did not require extracellular calcium, was sufficient to initiate the events leading to increased TH expression. Consistent with this, the effects of bradykinin on TH expression were inhibited by 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid or 3,4,5-trimethoxybenzoic acid 8-(diethylamino)-octyl ester which chelates or inhibits the release of intracellular calcium, respectively. Bradykinin required a rise in [Ca2+]i for <10 min, as opposed to 10–30 min for depolarization to increase TH mRNA levels. These results demonstrate that although each of these treatments increased TH gene expression by raising [Ca2+]i, there are important differences among them in terms of the magnitude of elevated [Ca2+]i, requirements for extracellular calcium or release of intracellular calcium stores, and duration of elevated [Ca2+]i, indicating the involvement of different calcium signaling pathways leading to regulation of TH gene expression. 相似文献
62.
An analysis of the effects of inbreeding on the genetic structure of a colonizing population of Drosophila subobscura has been carried out. Species of Drosophila, particularly D. subobscura, may have lethal alleles associated with chromosomal inversions and our aim was to assess the extent to which the genome is balanced in this way. The frequencies of chromosomal inversions were compared between a large population and a set of 72 lines that were maintained by brother-sister mating for 10 generations. Fisher's matrix method was used to calculate the expected homozygosity in these inbred lines for 5 allozyme loci (Aph, Hk-1, Lap, Odh, and Pept-1) used as markers of large chromosomal segments. Furthermore, the expected rates of fixation corresponding to these allozyme loci were also calculated. The results show that the amount of homozygosis observed did not differ significantly from expectations (with the corresponding loss of lines as a consequence of the reduction in viability). However, two deviations from strict neutrality were observed: there was a heterozygote excess at the Lap locus, and the frequency of the O
5 inversion (always associated with a lethal gene in colonizing populations) was higher than expected. 相似文献
63.
DNA Segments Sensitive to Single-Strand-Specific Nucleases Are Present in Chromatin of Mitotic Cells
It was observed before that DNAin situin chromatin of mitotic cells is more sensitive to denaturation than DNA in chromatin of interphase cells. DNA sensitivity to denaturation, in these studies, was analyzed by exposing cells to heat or acid and using acridine orange (AO), the metachromatic fluorochrome which can differentially stain double-stranded (ds) vs single-stranded (ss) nucleic acids, as a marker of the degree of DNA denaturation. However, without prior cell treatment with heat or acid no presence of single-stranded DNA in either mitotic or interphase cells was detected by this assay. In the present experiments we demonstrate that DNAin situin mitotic cells, without any prior treatment that can induce DNA denaturation, is sensitive to ss-specific S1 and mung bean nucleases. Incubation of permeabilized human T cell leukemic MOLT-4, promyelocytic HL-60, histiomonocytic lymphoma U937 cells, or normal PHA-stimulated lymphocytes with S1 or mung bean nucleases generated extensive DNA breakage in mitotic cells. DNA strand breaks were detected using fluorochrome-labeled triphosphonucleotides in the reaction catalyzed by exogenous terminal deoxynucleotidyl transferase. Under identical conditions of the cells’ exposure to ss-specific nucleases, DNA breakage in interphase cells was of an order of magnitude less extensive compared to mitotic cells. The data indicate that segments of DNA in mitotic chromosomes, in contrast to interphase cells, may be in a conformation which is sensitive to ss nucleases. This may be a reflection of the differences in the torsional stress of DNA loops between interphase and mitotic chromatin. Namely, greater stress in mitotic loops may lead to formation of the hairpin-loop structures by inverted repeats; such structures are sensitive to ss nucleases. The present method of detection of such segments appears to be more sensitive than the use of AO. The identification of mitotic cells based on sensitivity of their DNA to ss nucleases provides an additional method for their quantification by flow cytometry. 相似文献
64.
Márcia Camargo-De-Morais Marta De Freitas Angela G. De Mattos Nádia Schröder Ana C. Zilles Carla S. F. Lisboa Nice Arteni Armando Barlem Rejane Schierholt Guilherme Zwetsch Carlos A. B. Souza Regina Pessoa-Pureur Carlos A. Netto 《Neurochemical research》1996,21(5):595-602
Neurofilaments subunits (NF-H, NF-M, NF-L) and glial fibrillary acidic protein (GFAP) were investigated in the hippocampus
of rats after distinct periods of reperfusion (1 to 15 days) following 20 min of transient global forebrain ischemia in the
rat. In vitro [14Ca]leucine incorporation was not altered until 48 h after the ischemic insult, however concentration of intermediate filament
subunits significantly decreased in this period. Three days after the insult, leucine incorporation significantly increased
while the concentration NF-H, NF-M, and NF-L were still diminished after 15 days of reperfusion. In vitro incorporation of32P into NF-M and NF-L suffered immediately after ischemia, but returned to control values after two days of reperfusion. GFAP
levels decreased immediately after ischemia but quickly recovered and significantly peaked from 7 to 10 days after the insult.
These results suggest that transient ischemia followed by reperfusion causes proteolysis of intermediate filaments in the
hippocampus, and that proteolysis could be facilitated by diminished phosphorylation levels of NF-M and NF-L. 相似文献
65.
Ana Maria Landeira-Fernandez Maricilia S. Costa Leopolde de Meis 《Bioscience reports》1996,16(6):439-451
Vesicles derived from maize roots retain a membrane bound H+-ATPase that is able to pump H+ at the expense of ATP hydrolysis. In this work it is shown that heparin, fucose-branched chondroitin sulfate and dextran sulfate 8000 promote a shift of the H+-ATPase optimum pH from 6.0 to 7.0. This shift is a result of a dual effect of the sulfated polysaccharides, inhibition at pH 6.0 and activation at pH 7.O. At pH 6.0 dextran 8000 promotes an increase of the apparent Km for ATP from 0.28 to 0.95 mM and a decrease of the Vmax from 14.5 to 7.1 mol Pi/mg · 30 min–1. At pH 7.0 dextran 8000 promotes an increase in Vmax from 6.7 to 11.7 mol Pi/mg · 30 min–1. In the presence of lysophosphatidylcholine the inhibitory effect of the sulfated polysaccharides observed at pH 6.0 was not altered but the activation of pH 7.0 decreased. It was found that in the presence of sulfated polysaccharides the ATPase became highly sensitive to K+ and Na+. Both the inhibition at pH 6.0 and the activation promoted by the polysaccharide were antagonized by monovalent cations (K+>Na+Li+).Abbreviations Mops
4-morpholinopropanesulfonic acid
- EDTA
ethylenediaminetetraacetic acid
- ACMA
9-amino-6-chloro-2-methoxyacridine
- FCCP
carbonyl cyanide p(trifluoromethoxy)-phenylhyrazone 相似文献
66.
Results are presented on the phytoplankton species compositionand abundance from bottle samples collected in September 1989near the confluence of the Brazil and Malvinas currents offArgentina. The phytoplankton assemblages were dominated by diatomsand dinoflagellates. A surface diatom bloom was found alongthe west side of the Brazil Current, and was dominated by Thalassiosiradelicatula Ostenfeld emend. Hasle (cell numbers up to 5.5 x105 cells 11) The bloom was associated with strong temperaturegradients separating Brazil and Malvinas waters, and with thepresence of a cyclonic eddy near the confluence of the currents.These features were detected in satellite imagery coincidentwith the in situ sampling dates. 相似文献
67.
Rossella Serra Gloria Isani Otello Cattani Emilio Carpené 《Biological trace element research》1996,51(1):107-116
Gilthead were fed three diets. Diet A was the control diet, whereas diets B and C were supplemented with 300 and 900 mg Zn/kg,
respectively. Fish fed with diet C, at the end of the experiment, showed the lowest weight. Zinc concentrations presented
the higher values in gills, liver, and kidney. Muscle and brain had the lower mean values and showed a tight control of zinc
levels. These results reinforce the hypothesis that zinc in the CNS should be strictly controlled in order to maintain the
functional role of the metal. Significant differences in tissue zinc concentrations were obtained between fish fed different
amounts of zinc, the metal concentrations being higher in tissues of fish fed diet C. The tissue decrease of zinc, found at
the end of the experiment, may depend on a lower feed consumption or on different zinc requirements during the cold season.
These changes, even if not univocal among the three diets, may be associated with the life cycle of fish. Furthermore, copper
concentrations were little affected by the different concentrations of zinc in the three diets; liver and kidney presented
the highest concentrations; liver showed a significant decrease in copper content at the end of the experiment. We conclude
that: zinc concentrations of the diet may affect the gilthead weights and the tissual metal content; and zinc concentrations
in the diets, depending on the growth rate, may be varied depending on the season. 相似文献
68.
Ana Cristina Gomes da Cunha Manuel Fernandes Ferreira 《Plant Cell, Tissue and Organ Culture》1996,47(1):1-8
The effects of plant growth regulators (PGR) on calli induction, morphogenesis and somatic embryogenesis of flax were studied. The organogenic and callus formation capacity were assessed for different types of source explants. Root and shoot explants were equally good material for calli production but the former produced calli without shoot regeneration capacity. Under the experimental conditions tested, 2,4-dichlorophenoxyacetic acid (2,4-D) + zeatin was the most efficient PGR combination on calli induction and biomass production. The calli were green but with no rhizogenic capacity. In contrast, and at similar concentrations, indole-3-butyric acid (IBA) + kinetin induced white or pale green friable calli with a good root regeneration capacity (60%). A factorial experiment with different combinations of 2,4-D + zeatin + gibberellic acid (GA3) levels revealed that the direction of explant differentiation was determined by specific PGR interactions and concentrations. The results from these experiments revealed that the morphogenetic pathway (shoot versus root differentiation) can be manipulated on flax explants by raising the 2,4-D level from 0.05 to 3.2 mg l?1 in the induction medium. The induction and development of somatic embryos from flax explants was possible in a range of 2,4-D + zeatin concentrations surrounding 0.4 mg l?1 2,4-D and 1.6 mg l?1 zeatin, the most efficient growth regulator combination. 相似文献
69.
Ana María Planchuelo 《Brittonia》1996,48(2):263-265
Lupinus rubriflorus Planchuelo, from Rio Grande do Sul, Brazil, is described and illustrated. This species is related toL. bracteolaris s.l. differing in pubescence, stipules, and floral characters. 相似文献
70.
Sandra Irene Pitta-Alvarez Ana María Giulietti 《In vitro cellular & developmental biology. Plant》1995,31(4):215-220
Summary
Brugmansia candida hairy roots, obtained by infection withAgrobacterium rhizogenes LBA 9402, exhibit, after subculturing in liquid media, a tendency towards dedifferentiation. It has been found that the following
strategies can be applied to inhibit this dedifferentiation and preserve normal root morphology: (a) lowering both the mineral
and sucrose concentration in the media employed so as to diminish osmotic stress (a condition to which these roots appear
to be particularly susceptible); (b) employing antiauxins in appropriate concentrations; and (c) maintaining the hairy roots
on solid media prior to use in production processes in liquid media. The first strategy suggested does not favor alkaloid
productivity, but in this case a two-step method could be attempted: biomass with normal root morphology could be obtained
in a first stage using low sucrose concentrations, and in a second stage, sucrose could be increased in order to achieve higher
productivity. In all the clones ofB. candida obtained, alkaloid production was biased towards scopolamine. 相似文献