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21.
Paul G. Layer Regina Alber Patrick Mansky Günter Vollmer Elmar Willbold 《Cell and tissue research》1990,259(2):187-198
Summary We report here that similar to E6-chicken retinal cells, dissociated cells from 5.5-day-old (E5.5) quail retinae reaggregate in rotary culture, multiply about tenfold and reestablish histotypical areas. These cellular aggregates include all nuclear layers either with inversed or correct laminar polarity, depending on the local origin of the cells (called rosetted and laminar in-vitro-retinae (IVR), respectively; Layer and Willbold 1989). In combined cultures, chick and quail cells are evenly mixed only during the first two days of culture. Along with the assembly of single cells into rosettes and then into discrete laminae, sectors of chick and quail cells begin to segregate. They are delineated by borders running radially through all three nuclear layers. Thus, interspecies migration of cells at this advanced stage of differentiation is strongly inhibited. Concomitant with this segregation, coherent radial columns spanning all three layers but containing cells from either species only, can be traced histologically. We conclude that a weak segregation of chick and quail retinal cells takes place already at the single cell level, but that the permanent segregation of entire tissue parts must be due to clonal cellular proliferation within the IVR in conjunction with some developmental-structural mechanism retaining clonal progenies within a columnar order.Abbreviations
ECM
extracellular matrix
-
E5.5
days of embryonic age
-
GCL
ganglion cell layer
-
GC's
ganglion cells
-
i.c.
in culture
-
INL
inner nuclear layer
- rosetted in-vitro-retina
retinal cell organoid aggregated from single cells of the central retina
-
IPL
inner plexiform layer
-
MRE
marginal retinal epithelium
-
ONL
outer nuclear layer
-
OPL
outer plexiform layer
-
OS
ora serrate
-
PR
photoreceptor cell
- laminated in-vitro-retina
fully laminated retinal cellorganoid resembling an E15-retina aggregated from cells of the eye periphery including RPE
-
RPE
retinal pigment epithelium 相似文献
22.
Influence of Calcium Ion on Ethanol Tolerance of Saccharomyces bayanus and Alcoholic Fermentation by Yeasts 总被引:1,自引:0,他引:1 下载免费PDF全文
Regina C. Nabais Isabel S-Correia Cristina A. Viegas Júlio M. Novais 《Applied microbiology》1988,54(10):2439-2446
The addition of Ca2+ (as CaCl2) in optimal concentrations (0.75 to 2.0 mM) to a fermentation medium with a trace contaminating concentration of Ca2+ (0.025 mM) led to the rapid production of higher concentrations of ethanol by Saccharomyces cerevisiae, Saccharomyces bayanus, and Kluyveromyces marxianus. The positive effect of calcium supplementation (0.75 mM) on alcoholic fermentation by S. bayanus was explained by the increase in its ethanol tolerance. The ethanol inhibition of growth and fermentation followed the equation μxi = μoi [1 - (X/Xmi)]ni, where μoi and μxi are, respectively, the specific growth (i = g) and fermentation (i = f) rates in the absence or presence of a concentration (X) of added ethanol, and Xmi is the maximal concentration of ethanol which allows growth or fermentation. The toxic power is given by ni. In Ca2+ - supplemented medium (0.75 mM), ng = 0.42 for growth and nf = 0.43 for fermentation compared with 0.52 and 0.55, respectively, in unsupplemented medium; for both media, Xmg = 10% (vol/vol) and Xmf = 13% (vol/vol). For lethal concentrations of ethanol, the specific death rates were minimal for cells that were grown and incubated with ethanol in medium with an optimal concentration of Ca2+, maximal for cells grown and incubated with ethanol in unsupplemented medium, and intermediate for cells grown in unsupplemented medium and incubated with ethanol in calcium-supplemented medium. The effect of Ca2+ on the acidification curve of energized cells in the presence of ethanol was found to be closely associated with its protective effect on growth, fermentation, and viability. 相似文献
23.
Many studies have established a correlation of differences in the activities of various muscle types with differences in the expression of myosin isoforms. In this paper we report the sequence determination of myosin light chain-2 from rabbit slow skeletal (LC2s) and ventricular (LC2v) nmscles. We sequenced tryptic peptides from LC2v which account for all except a few terminal amino acid residues. The major part (87 residues) of the rabbit LC2s sequence, obtained from tryptic and cyanogen bromide (CNBr) peptides, was found to be identical to rabbit LC2v. Our results provide the first sequence information on LC2s from any species, and lend strong support to the hypothesis that LC2s and LC2v are identical. Comparisons of rabbit LC2v and LC2s with rabbit LC2f (from fast skeletal muscle), and also with chicken LC2f and LC2v, show clearly that LC2s and LC2v from mammalian and avian species are more closely related to each other than they are to LC2f isoforms from the same species. 相似文献
24.
25.
G. Menestrina C. Pederzolli M. Dalla Serra M. Bregante F. Gambale 《The Journal of membrane biology》1996,149(2):113-121
Escherichia coli hemolysin is known to cause hemolysis of red blood cells by forming hydrophilic pores in their cell membrane. Hemolysin-induced
pores have been directly visualized in model systems such as planar lipid membranes and unilamellar vesicles. However this
hemolysin, like all the members of a related family of toxins called Repeat Toxins, is a potent leukotoxin. To investigate
whether the formation of channels is involved also in its leukotoxic activity, we used patch-clamped human macrophages as
targets. Indeed, when exposed to the hemolysin, these cells developed additional pores into their membrane. Such exogenous
pores had properties very different from the endogenous channels already present in the cell membrane (primarily K+ channels), but very similar to the pores formed by the toxin in purely lipidic model membranes. Observed properties were:
large single channel conductance, cation over anion selectivity but weak discrimination among different cations, quasilinear
current-voltage characteristic and the existence of a flickering pre-open state of small conductance. The selectivity properties
of the toxin channels appearing in phospholipid vesicles were also investigated, using a specially adapted polarization/depolarization
assay, and were found to be completely consistent with that of the current fluctuations observed in excised macrophage patches.
Received: 14 August 1995/Revised: 2 October 1995 相似文献
26.
Birgit Conrad Regina S. Savchenko Roland Breves Jürgen Hofemeister 《Molecular & general genetics : MGG》1996,250(2):230-236
The adaptation and application of theEscherichia coli T7 RNA polymerase system for regulated and promoter-specific gene expression inBacillus subtilis is reported. The expression cassette used inBacillus subtilis was tightly regulated and T7 RNA polymerase (T7 RNAP) appeared 30 min after induction. The efficiency of T7 promoter-specific gene expression inB. subtilis was studied using one secretory and two cytosolic proteins of heterologous origin. The accumulation ofE. coli -galactosidase, as well as a 1,4--glucosidase fromThermoanaerobacter brockii inB. subtilis after T7 RNAP induction was strongly enhanced by rifampicin inhibition of host RNAP activity. The-amylase ofThermoactinomyces vulgaris, a secretory protein, was found to accumulate in the culture supernatant up to levels of about 70 mg/l 10–20 h after T7 RNAP induction, but was also deposited in cellular fractions. The addition of rifampicin inhibited-amylase secretion, but unexpectedly, after a short period, also prevented its further (intra)cellular accumulation 相似文献
27.
Luciana A. Haddad Regina C. Mingroni-Netto Angela M. Vianna-Morgante Sérgio D. J. Pena 《Human genetics》1996,97(6):808-812
Ever since the identification of the genetic cause of fragile X syndrome as the expansion of an unstable trinucleotide sequence,
several diagnostic strategies have evolved from molecular studies. However, we still lack a simple test suitable for population
screening. We have therefore developed a nonisotopic polymerase chain reaction (PCR)-based technique for the identification
of fragile X full mutations among men, with easy visualization of the PCR products on silver-stained polyacrylamide gels.
The technique consists of PCR amplification with primers that flank the trinucleotide repeats, with a product of 557 bp for
the (CGG)29 allele. Conditions were established such that full mutations failed to amplify and were thus identified with 98% sensitivity
compared with Southern blot analysis. To produce an indispensable internal control we added to the reaction a third primer,
internal to this fragment, allowing the multiplex amplification of a monomorphic band corresponding to a CG-rich stretch 147
bp upstream of the polymorphic region. In trials involving 41 patients and 74 controls, the PCR-based test here described
showed specificity of more than 98.6%, accuracy of 99% and a sensitivity of 98%. Thus, although not suitable for medical diagnosis,
it constitutes a useful tool for screening for the fragile X syndrome in populations of mentally retarded males.
Received: 31 May 1995 / Revised: 4 October 1995 相似文献
28.
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
29.
The human c-erbB-2 oncogene is homologous to the ratneu oncogene, both encoding transmembrane growth factor receptors. Overexpression and point mutations in the transmembrane domain of the encoded proteins in both cases have been implicated in cell transformation and carcinogenesis. In the case of theneu protein, it has been proposed that these effects are mediated by conformational preferences for anα-helix in the transmembrane domain, which facilitates receptor dimerization, an important step in the signal transduction process. To examine whether this is the case for c-erbB-2 as well, we have used conformational energy analysis to determine the preferred three-dimensional structures for the transmembrane domain of the c-erbB-2 protein from residues 650 to 668 with Val (nontransforming) and Glu (transforming) at position 659. The global minimum energy conformation for the Val-659 peptide from the normal, nontransforming protein was found to contain several bends, whereas the global minimum energy conformation for Glu-659 peptide from the mutant, transforming protein was found to beα-helical. Thus, the difference in conformational preferences for these transmembrane domains may explain the difference in transforming ability of these proteins. The presence of higher-energyα-helical conformations for the transmembrane domain from the normal Val-659 protein may provide an explanation for the presence of a transforming effect from overexpression of c-erbB-2. In addition, docking of the oncogenic sequences in theirα-helical and bend conformations shows that the all-α-helical dimer is clearly favored energetically over the bend dimer. 相似文献
30.
Effect of tumor necrosis factor-α and interferon-γ on the growth of a human salivary gland cell line
Ava J. Wu Regina H. Kurrasch Joseph Katz Philip C. Fox Bruce J. Baum Jane C. Atkinson 《Journal of cellular physiology》1994,161(2):217-226
Interferon-γ (IFN-γ) is a product of activated T-lymphocytes, and tumor necrosis factor-α (TNF-α) is a product of both lymphocytes and macrophages. These cell types are often present at sites of tissue damage secondary to chronic infection or autoimmune disease. The purpose of this study was to characterize the effects of TNF-α and IFN-γ on a human submandibular gland epithelial cell line (HSG). IFN-γ caused a concentration-dependent decrease in HSG cell growth (~70% in 6 days). Conversely, TNF-α alone had little effect on the growth of these cells. When these cytokines were added in combination (20 units/ml TNF-α and 1,000 units/ml of IFN-γ), there was a synergistic antiproliferative effect; no apparent cell growth was observed. The cytokine-induced antiproliferative effect was reversible. After the apparent cessation of cell growth for 3–6 days, removal of the cytokines permitted complete growth recovery. Further, cells that recovered and exhibited growth patterns that were similar to control cells remained susceptible to the antiproliferative effects of the cytokines. Flow cytometry revealed that the percentage of cells in G0/G1 with the combination of cytokines was significantly increased by 24 h. The antiproliferative effect of IFN-γ alone and that of IFN-γ and TNF-α in combination were blocked completely using an antibody to the IFN-γ receptor. A hypothesized mechanism of tissue damage in autoimmune inflammatory disorders is via up-regulation of cell surface markers such as intercellular adhesion molecule type I (ICAM-1) and histocompatibility antigen HLA-DR which can exacerbate the inflammatory process. Treatment of HSG cells with IFN-γ, with or without TNF-α, resulted in increased levels of ICAM-1 and the acquisition of HLA-DR expression. These aggregate data suggest that IFN-γ alone can regulate the expression of cell surface markers involved in the inflammatory process as well as cause a potent yet reversible inhibition of HSG cell growth that is modulated by the presence of TNF-α. © 1994 Wiley-Liss, Inc. 1 This article is a US Government work and, as such, is in the public domain in the United States of America. 相似文献