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41.
Hilde Beele Hubert Thierens Leo de Ridder 《In vitro cellular & developmental biology. Plant》1989,25(10):923-933
Summary Different organotypical culture methods are used to test the direct effects of serotonin and ketanserin, a S2, α1, and H1 receptor antagonist in vascular tissue, on fibroblasts and epidermal cells of embryonic chick skin in vitro. From light microscopic
and electron microscopic analyses, we learn that serotonin enhances keratinization and differentiation, whereas ketanserin
reduces differentiation in comparison to the control cultures. Incorporation data of fragments cultured with [3H]thymidine show that ketanserin, within a dose range from 0.05 to 5 μg/ml, stimulates proliferation. Serotonin at a concentration
of 10 μg/ml slightly slows down proliferation, whereas lower doses of 0.1 and 1 μg/ml result in tritium activities that do
not differ from control cultures.
This investigation was financially supported by the National Fund of Scientific Research, Belgium, 3.0022.87. 相似文献
42.
Hubert Wiener Dan A. Klaerke Peter L. Jørgensen 《The Journal of membrane biology》1990,117(3):275-283
Summary In the mammalian distal colon, the surface epithelium is responsible for electrolyte absorption, while the crypts are the site of secretion. This study examines the properties of electrical potential-driven86Rb+ fluxes through K+ channels in basolateral membrane vesicles of surface and crypt cells of the rabbit distal colon epithelium. We show that Ba2+-sensitive, Ca2+-activated K+ channels are present in both surface and crypt cell derived vesicles with half-maximal activation at 5×10–7
m free Ca2+. This suggests an important role of cytoplasmic Ca2+ in the regulation of the bidirectional ion fluxes in the colon epithelium.The properties of K+ channels in the surface cell membrane fraction differ from those of the channels in the crypt cell derived membranes. The peptide toxin apamin inhibits Ca2+-activated K+ channels exclusively in surface cell vesicles, while charybdotoxin inhibits predominantely in the crypt cell membrane fraction. Titrations with H+ and tetraethylammonium show that both high-and low-sensitive86Rb+ flux components are present in surface cell vesicles, while the high-sensitive component is absent in the crypt cell membrane fraction. The Ba2+-sensitive, Ca2+-activated K+ channels can be solubilized in CHAPS and reconstituted into phospholipid vesicles. This is an essential step for further characterization of channel properties and for identification of the channel proteins in purification procedures. 相似文献
43.
Pharmacological Characterization of Somatostatin Receptors in the Rat Cerebellum During Development 总被引:5,自引:2,他引:3
Bruno J. Gonzalez Philippe Leroux Corinne Bodenant Pierre Braquet Hubert Vaudry 《Journal of neurochemistry》1990,55(3):729-737
Abstract: Somatostatin (SRIF) receptors (SRIF-Rs) are transiently expressed in a germinative lamina of the rat cerebellum, the external granule cell layer. The appearance of SRIF-Rs coincides with the expression of SRIF-like immunoreactivity in the cerebellum. However, the cellular location of SRIF-Rs does not overlap with the distribution of SRIF-like immunoreactivity, with the latter being restricted to ascending fibers arising from the brainstem, to perikarya within the white matter, and to some Purkinje cells. The characterization of SRIF-Rs in the immature (13–day-old) rat cerebellum was conducted by means of binding experiments in membraneenriched preparations and autoradiography, using two radioligands, [125I-Tyr0,D-Trp8]SRIF-14 ([125I-Tyr0,d -Trp8]S14) and I25I-SMS 204–090. The pharmacological profile of cerebellar SRIF-Rs was compared with that of adult cortical SRIF-Rs. Saturation studies performed in 13–day-old rat cerebellum showed that the A'D values for [125I-Tyr0,D-Trp8]S14 and 125I-SMS 204–090 binding were 0.35 ± 0.04 and 0.39 ± 0.01 nM, respectively. The corresponding Bmax values were 52.7 ± 4.8 and 49.9 ± 5.3 fmol/mg of protein, a result indicating that radioligands with high specific radioactivity (2,000 Ci/mmol) bind to a single class of high-affinity sites (SSI). Competition studies showed that different D-Trp-sub-stituted analogs displaced [125I-Tyr0,d -Trp8]S14 binding with Hill coefficients >1, a finding indicating the existence of different subtypes of binding sites. When [Tyr0,d -Trp8]S14 was used as a competitor, two sites were resolved by Scatchard analysis in both 13–day-old cerebellum and adult cerebral cortex. The higher-affinity sites correspond to the SSI subtype identified in saturation experiments, whereas the lower-affinity sites most likely correspond to the SS2 subtype. Ionic supplementation studies showed that divalent cations were required to obtain maximal specific binding on the SSI sites. In particular, Mn2+ was the most efficient cation for promoting binding of [125I-Tyr0,d -Trp8]S14. Addition of GTP to the incubation buffer induced a marked reduction of specific binding. The results obtained by membrane binding assays were similar to those obtained by quantitative autoradiography, a result indicating that the microenvironment of SRIF-Rs was preserved in both types of tissue preparations. Receptors expressed in the developing rat cerebellum exhibited the same KD and similar pharmacological profile as those observed in the adult rat cortex. These results show that SRIF-binding sites transiently expressed in the external granule cell layer of the cerebellum of young rats are indistinguishable from adult rat brain SRIF-Rs. The extremely high density of SRIF-Rs found in the external granule cell layer in 13–day-old rats suggests that SRIF may play a pivotal role in the proliferation and/or differentiation of these germinative cells. 相似文献
44.
Peter Raeymaekers Christine Van Broeckhoven Hubert Backhovens Anita Wehnert Ludo Muylle Peter De Jonghe Jan Ghenens Jean-Jacques Martin Antoon Vandenberghe 《Human genetics》1989,81(3):231-233
Summary We have recently demonstrated tight linkage of the Duffy blood group marker to the -spectrin gene in an extended pedigree with Charcot-Marie-Tooth neuropathy. To determine a more precise location of the Duffy blood group locus on the chromosome 1 map we have tested several more chromosome 1 genes for linkage with this marker. We found suggestive linkage with the antithrombin III and apolipoprotein A2 genes and conclusive linkage with the gene coding for -nerve growth factor. 相似文献
45.
Claude Penel Thomas Gaspar Michèle Crèvecoeur Claire Kevers Hubert Greppin 《Physiologia plantarum》1990,79(2):250-254
Ca2+ and Mn2+ activate the conversion of 1-aminocyclopropane-1-carboxylic acid (ACC) by root microsomes of Vicia lens as they do in other similar systems. The preparation of microsomes in the presence of Mn2+ greatly increases their ability to convert ACC into ethylene, without addition of Mn2+ in the reaction mixture. Ca2+ does not have this property. The effect could not be attributed to Mn2+ entrapping into membrane vesicles (sonication followed by repelleting had no effect) but, possibly, in part to Mn2+ -mediated binding to microsomes of a soluble factor favouring the conversion of ACC to C2 H4 . Although no direct correlation could be established in vitro between ethylene-forming-enzyme (EFE) and peroxidase activities, some soluble peroxidases might be this soluble factor. Mn2+ favoured attachment to membranes of some peroxidase activity from the soluble fraction and from commercial HRP and lipoxygenase. This binding effect of Mn2+ cannot be readily distinguished from its role in the generation of a chain of free radicals and in redox mechanisms. 相似文献
46.
In Vivo and In Vitro Evidence for the Biosynthesis of Testosterone in the Telencephalon of the Female Frog 总被引:5,自引:2,他引:3
Ayikoe G. Mensah-Nyagan Jean-Luc Do-Rego Marc Feuilloley Albert Marcual Catherine Lange †Georges Pelletier Hubert Vaudry 《Journal of neurochemistry》1996,67(1):413-422
Abstract: Neurons and glial cells are capable of synthesizing various steroid hormones, but biosynthesis of testosterone in the CNS has never been reported. The aim of the present study was to demonstrate the synthesis of testosterone in the frog brain. The presence of 17β-hydroxysteroid dehydrogenase (17β-HSD)-like immunoreactivity was detected in a population of glial cells located in the telencephalon. Reversed-phase HPLC analysis of brain tissue extracts combined with radioimmunoassay detection revealed the presence of substantial amounts of testosterone and 5α-dihydrotestosterone (5α-DHT) in the telencephalon where 17β-HSD-positive cells were visualized. In male frogs, castration totally suppressed testosterone and 5α-DHT in the blood and in the rhombencephalon but did not affect the concentration of these two steroids in the telencephalon. Chemical characterization of testosterone in female frog telencephalon extracts was performed by coupling HPLC analysis with gas chromatography-mass spectrometry. Using the pulse-chase technique with [3 H]pregnenolone as a precursor, the formation of a series of metabolites was observed, including dehydroepiandrosterone, androstenedione, testosterone, 5α-DHT, and estradiol. These data demonstrate the existence of an active form of 17β-HSD in the frog telencephalon, which is likely involved in testosterone biosynthesis within the brain. 相似文献
47.
Olivier Cohen Christine Cans Jean Louis Gilardi Hubert Roth Marie-Ange Mermet Pierre Jalbert Jacques Demongeot Martine Cuillel 《Human genetics》1996,97(5):659-667
Reciprocal translocations (rcp) are among the most common constitutional chromosomal aberrations in man. Using a European
database of 1574 families carrying autosomal rcp, a cartographic study was done on the breakpoints involved. The breakpoints
are non-randomly distributed along the different chromosomes, indicating “hot spots”. Breakpoints of rcp that result in descendants
that are unbalanced chromosomally at birth are more frequent in a distal position on chromosomal arms, and 65% of them are
localised in R-bands. Among the R-bands, bands rich in GC islands and poor in Alu repetitive sequences are more frequently
the site of breakpoints, as well as bands that include a fragile site. This result suggests that the variation in degree of
methylation in GC islands could be involved in chromosomal breakage and hence in chromosomal rearrangements.
Received: 10 April 1995 / Revised: 1 July 1995 相似文献
48.
J.Michael Conlon Nicolas Chartrel Jerome Leprince Charles Suaudeau Jean Costentin Hubert Vaudry 《Peptides》1996,17(8):1291-1296
A peptide derived from the posttranslational processing of proenkephalin A was isolated from an extract of the brain of the European green frog Rana ridibunda and its primary structure established as: Tyr-Gly-Gly-Phe-Met-Arg-Arg-Val-Gly-Arg10-Pro-Glu-Trp-Trp-Gln-Asp-Tyr-Gln-Lys-Arg20-Tyr-Gly-Gly-Phe-Met. The structure was confirmed by chemical synthesis. The peptide represents an amphibian equivalent of bovine adrenal peptide E [preproenkephalin A (206–230)-peptide] but the sequence contains two amino acid substitutions (Met15 → Gln and Leu25 → Met) compared with the mammalian peptide. The data support previous hypotheses that the Leu-enkephalin sequence is not present in preproenkephalin A of amphibians. Intracerebroventricular injections of frog peptide E (10 and 100 ng) in mice had no significant effect on horizontal locomotor activity. The peptide, in doses up to 1 μg, had no effect on latency of escape jumping in the hot plate test and the peptide (100 ng) did not modify responses (paw licking, rearing, and escape jumping) in morphine-treated mice. 相似文献
49.
In the short day plant Chenopodium rubrum and the long day plant Nicotiana tabacum cv. Havana 425, adenylate kinase (EC 2.7.4.3) occurs as a family of isoforms, with at least two members localized in the chloroplast representing the main isoforms. In this work, isoforms were separated by anion exchange chromatography and relative isoform activities were compared between vegetative plants and plants induced to flowering. In both species examined, a light regime leading to floral induction resulted in a significant decrease in the activity of one chloroplast isoform. This decrease modified considerably the relative distribution of isoform activities, especially that between the two chloroplast activities. 相似文献
50.
A protein binds the selenocysteine insertion element in the 3'-UTR of mammalian selenoprotein mRNAs. 总被引:6,自引:0,他引:6
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Several gene products are involved in co-translational insertion of selenocysteine by the tRNA(Sec). In addition, a stem-loop structure in the mRNAs coding for selenoproteins is essential to mediate the selection of the proper selenocysteine UGA codon. Interestingly, in eukaryotic selenoprotein mRNAs, this stem-loop structure, the selenocysteine insertion sequence (SECIS) element, resides in the 3'-untranslated region, far downstream of the UGA codon. In view of unravelling the underlying complex mechanism, we have attempted to detect RNA-binding proteins with specificity for the SECIS element. Using mobility shift assays, we could show that a protein, present in different types of mammalian cell extracts, possesses the capacity of binding the SECIS element of the selenoprotein glutathione peroxidase (GPx) mRNA. We have termed this protein SBP, for Secis Binding Protein. Competition experiments attested that the binding is highly specific and UV cross-linking indicated that the protein has an apparent molecular weight in the range of 60-65 kDa. Finally, some data suggest that the SECIS elements in the mRNAs of GPx and another selenoprotein, type I iodothyronine 5' deiodinase, recognize the same SBP protein. This constitutes the first report of the existence of a 3' UTR binding protein possibly involved in the eukaryotic selenocysteine insertion mechanism. 相似文献