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41.
The presence of an Na+/Ca2+ exchange system in basolateral plasma membranes from rat small intestinal epithelium has been demonstrated by studying Na+ gradient-dependent Ca2+ uptake and the inhibition of ATP-dependent Ca2+ accumulation by Na+. The presence of 75 mM Na+ in the uptake solution reduces ATP-dependent Ca2+ transport by 45%, despite the fact that Na+ does not affect Ca2+-ATPase activity. Preincubation of the membrane vesicles with ouabain or monensin reduces the Na+ inhibition of ATP-dependent Ca2+ uptake to 20%, apparently by preventing accumulation of Na+ in the vesicles realized by the Na+-pump. It was concluded that high intravesicular Na+ competes with Ca2+ for intravesicular Ca2+ binding sites. In the presence of ouabain, the inhibition of ATP-dependent Ca2+ transport shows a sigmoidal dependence on the Na+ concentration, suggesting cooperative interaction between counter transport of at least two sodium ions for one calcium ion. The apparent affinity for Na+ is between 15 and 20 mM. Uptake of Ca2+ in the absence of ATP can be enhanced by an Na+ gradient (Na+ inside > Na+ outside). This Na+ gradient-dependent Ca2+ uptake is further stimulated by an inside positive membrane potential but abolished by monensin. The apparent affinity for Ca2+ of this system is below 1 μM. In contrast to the ATP-dependent Ca2+ transport, there is no significant difference in Na+ gradient-dependent Ca2+ uptake between basolateral vesicles from duodenum, midjejunum and terminal ileum. In duodenum the activity of ATP-driven Ca2+ uptake is 5-times greater than the Na+/Ca2+ exchange capacity but in the ileum both systems are of equal potency. Furthermore, the Na+/Ca2+ exchange mechanism is not subject to regulation by 1α,25-dihydroxy vitamin D-3, since repletion of vitamin D-deficient rats with this seco-steroid hormone does not influence the Na+/Ca2+ exchange system while it doubles the ATP-driven Ca2+ pump activity. 相似文献
42.
Sulfation of fucoidin in focus embryos: III. Required for localization in the rhizoid wall 下载免费PDF全文
Zygotes of the brown alga Fucus distichus L. Powell accumulate a sulfated polysaccharide (fucoidin) in the cell wall at the site of rhizoid formation. Previous work indicated that zygotes grown in seawater minus sulfate do not sulfate the preformed fucan (an unsulfated fucoidin) but form rhizoids. Under these conditions, we determined whether sulfation of the fucan is required for its localization in the rhizoid wall. This was accomplished by developing a specific stain for both the fucan and fucoidin. Using a precipitin assay, we demonstrated in vitro that the lectin ricin (RCA(I)) specifically complexes with both the sulfated and desulfated polysaccharide. No precipitate is observed when either is incubated in 0.1 M D-galactose or when RCA(I) is mixed with laminarin or alginic acid, the other major polysaccharides in Fucus. RCA(I) conjugated with fluorescein isothiocyanate (FITC) is also shown to bind specifically to fucoidin using a filter paper (DE81) assay. When added to zygotes, RCA(I)-FITC binds only to the site of fucoidin localization, i.e., the rhizoid cell wall. However, RCA(I)-FITC is not observed in the rhizoid wall of zygotes grown in the absence of sulfate. This observation is not due to inability of RCA(I)-FITC to bind to the fucan in vivo. Chemically desulfated cell walls that contained fucoidin in the rhizoid wall bind RCA(I)-FITC only in the rhizoid region. Also, the concentration of fucose-containing polymers and polysaccharides that form precipitates with RCA(I) is the same in embryos grown in the presence or absence of sulfate. If sulfate is added back to cultures of zygotes grown without sulfate, fucoidin is detected at the rhizoid tip by RCA(I)-FITC several hours later. These results support the conclusion that the enzymatic sulfation of the fucan is a modification of the polysaccharide required for its localization and/or assembly into a specific region of the cell wall. 相似文献
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44.
Matthijs Verhage Elly Besselsen Fernando H. Lopes Da Silvas Wim E. J. M. Ghijsen 《Journal of neurochemistry》1989,53(4):1188-1194
In the present study, we have investigated the role of Ca2+ in the coupling of membrane depolarization to neurotransmitter secretion. We have measured (a) intracellular free Ca2+ concentration ([Ca2+]i) changes, (b) rapid 45Ca2+ uptake, and (c) Ca2+-dependent and -independent release of endogenous glutamate (Glu) and gamma-aminobutyric acid (GABA) as a function of stimulus intensity by elevating the extracellular [K+] to different levels in purified nerve terminals (synaptosomes) from rat hippocampus. During stimulation, Percoll-purified synaptosomes show an increased 45Ca2+ uptake, an elevated [Ca2+]i, and a Ca2+-dependent as well as a Ca2+-independent release of both Glu and GABA. With respect to both amino acids, synaptosomes respond on stimulation essentially in the same way, with maximally a fourfold increase in Ca2+-dependent (exocytotic) release. Ca2+-dependent transmitter release as well as [Ca2+]i elevations show maximal stimulation at moderate depolarizations (30 mM K+). A correlation exists between Ca2+-dependent release of both Glu and GABA and elevation of [Ca2+]i. Ca2+-dependent release is maximally stimulated with an elevation of [Ca2+]i of 60% above steady-state levels, corresponding with an intracellular concentration of approximately 400 nM, whereas elevations to 350 nM are ineffective in stimulating Ca2+-dependent release of both Glu and GABA. In contrast, Ca2+-independent release of both Glu and GABA shows roughly a linear rise with stimulus intensity up to 50 mM K+. 45Ca2+ uptake on stimulation also shows a continuous increase with stimulus intensity, although the relationship appears to be biphasic, with a plateau between 20 and 40 mM K+.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
45.
Matthijs Verhage Hugo Sandman Fred Mosselveld Marco van de Velde Pieter A. Hengst Fernando H. Lopes da Silva Wim E. J. M. Ghijsen 《Journal of neurochemistry》1992,58(4):1313-1320
To study the release of neurotransmitters, i.e., the recruitment of transmitters for release and the regulation of the release process, isolated nerve terminals (synaptosomes) of the rat forebrain were immobilized in Sephadex gel inside a perfusion chamber. In this way, the following were achieved: (a) A very limited pressure stress was exerted on the synaptosomes, so that these remained viable for long periods (greater than 30 min) inside the chamber and did not elute from the chamber, which allowed long-term experiments with repeated stimulations; (b) estimation of the release of various endogenous transmitters, both in a Ca(2+)-dependent (exocytotic) and Ca(2+)-independent manner; (c) a step-like stimulation with depolarizing agents (rise time, 3-4 s) and a high time resolution (600-ms sampling); and (d) negligible reuptake of transmitter into the terminals or extracellular breakdown. It is concluded that this perfusion setup helps to provide new insights in the presynaptic stimulus-secretion coupling, co-transmission, and the exo-endocytosis cycle. 相似文献
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47.
Calcium transport into brush border vesicles from rat small intestine was investigated by determining uptake rates at very short incubation periods. At incubation times up to 1 second a linear relationship between calcium uptake and time was observed at free calcium concentrations ranging from 1 microM to 5 mM. At time points above 1 second calcium uptake deviates progressively from linearity. Several lines of evidences (EGTA-wash, dependency on membrane potential, temperature sensitivity and effect of the calcium ionophore A23187) suggest transmembrane transport rather than extravesicular binding of calcium as being responsible for calcium uptake. Saturation experiments performed under initial linear and curvilinear uptake conditions show a saturable transport component in the mu molar and only a tendency to saturate in the molar concentration range. It is concluded that uptake values far from equilibrium are characteristic for transmembrane flux of calcium. Transmembrane flux of calcium is mediated by multiple and potential-sensitive mechanisms. 相似文献
48.
Lopez JV; Culver M; Stephens JC; Johnson WE; O'Brien SJ 《Molecular biology and evolution》1997,14(3):277-286
Differential rates of nucleotide substitution among different gene segments
and between distinct evolutionary lineages is well documented among
mitochondrial genes and is likely a consequence of locus-specific selective
constraints that delimit mutational divergence over evolutionary time. We
compared sequence variation of 18 homologous loci (15 coding genes and 3
parts of the control region) among 10 mammalian mitochondrial DNA genomes
which allowed us to describe different mitochondrial evolutionary patterns
and to produce an estimation of the relative order of gene divergence. The
relative rates of divergence of mitochondrial DNA genes in the family
Felidae were estimated by comparing their divergence from homologous
counterpart genes included in nuclear mitochondrial DNA (Numt, pronounced
"new might"), a genomic fossil that represents an ancient transfer of 7.9
kb of mitochondrial DNA to the nuclear genome of an ancestral species of
the domestic cat (Felis catus). Phylogenetic analyses of mitochondrial
(mtDNA) sequences with multiple outgroup species were conducted to date the
ancestral node common to the Numt and the cytoplasmic (Cymt) mtDNA genes
and to calibrate the rate of sequence divergence of mitochondrial genes
relative to nuclear homologous counterparts. By setting the fastest
substitution rate as strictly mutational, an empirical "selective
retardation index" is computed to quantify the sum of all constraints,
selective and otherwise, that limit sequence divergence of mitochondrial
gene sequences over time.
相似文献
49.
Loss of meiosis in Aspergillus 总被引:2,自引:0,他引:2
If strictly mitotic asexual fungi lack recombination, the conventional view
predicts that they are recent derivatives from older meiotic lineages. We
tested this by inferring phylogenetic relationships among closely related
meiotic and strictly mitotic taxa with Aspergillus conidial (mitotic)
states. Phylogenies were constructed by using DNA sequences from the
mitochondrial small ribosomal subunit, the nuclear ribosomal internal
transcribed spacers, and the nuclear 5.8S ribosomal gene. Over 920 bp of
sequence was analyzed for each taxon. Phylogenetic analysis of both the
mitochondrial and nuclear data sets showed at least four clades that
possess both meiotic and strictly mitotic taxa. These results support the
hypothesis that strictly mitotic lineages arise frequently from more
ancient meiotic lineages with Aspergillus conidial states. Many of the
strictly mitotic species examined retained characters that may be vestiges
of a meiotic state, including the production of sclerotia, sclerotium-like
structures, and hulle cells.
相似文献
50.
本文报道了石耳科的两个亚洲的,也是美洲以外的新记录种,即角石耳与深色石耳。进一步证实了单果石耳在南大西洋的,即南美洲和南部非洲的间断分布。对于上述种类以及本科其它一些种类的间断分布和替代现象进行了分析讨论。 相似文献