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  • Saccharomyces cerevisiae  (169)
  • Immunohistochemistry  (147)
  • Springer  (316)
  • American Institute of Physics
  • American Institute of Physics (AIP)
  • 2005-2009
  • 1985-1989  (316)
  • 1950-1954
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Year
  • 1
    Electronic Resource
    Electronic Resource
    Biotransformation of aromatic aldehydes bySaccharomyces cerevisiae: Investigation of reaction rates (1989)
    Springer
    Journal of industrial microbiology and biotechnology 4 (1989), S. 49-53 
    Details
    ISSN: 1476-5535
    Keywords: l-Phenylacetyl carbinol ; Saccharomyces cerevisiae ; Yeast ; Benzaldehyde ; Biotransformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The rate of production ofl-phenylacetyl carbinol bySaccharomyces cerevisiae in reaction mixtures containing benzaldehyde with sucrose or pyruvate as cosubstrate was investigated in short 1 h incubations. The effect of yeast dose rate, sucrose and benzaldehyde concentration and pH on the rate of reaction was determined. Maximum biotransformation rates were obtained with concentrations of benzaldehyde, sucrose and yeast of 6 g, 40 g and 60 g/l, respectively. Negligible biotransformation rates were observed at a concentration of 8 g/l benzaldehyde. The reaction had a pH optimum of 4.0–4.5. Rates of bioconversion of benzaldehyde and selected substituted aromatic aldehydes using both sucrose and sodium pyruvate as cosubstrate were compared. The rate of aromatic alcohol production was much higher when sucrose was used rather than pyruvate.o-Tolualdehyde and 1-chlorobenzaldehyde were poor substrates for aromatic carbinol formation although the latter produced significant aromatic alcohol in sucrose-containing media. Yields of 2.74 and 3.80 g/l phenylacetyl carbinol were produced from sucrose and pyruvate, respectively, in a 1 h reaction period.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569693
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  • 2
    Electronic Resource
    Electronic Resource
    High solids fermentation of hydrolysates of wheat starch B in a continuous dynamic immobilized biocatalyst bioreactor (1989)
    Springer
    Journal of industrial microbiology and biotechnology 4 (1989), S. 81-84 
    Details
    ISSN: 1476-5535
    Keywords: Ethanol fermentation ; Wheat starch ; Saccharomyces cerevisiae ; immobilization ; Continuous dynamic immobilized biocatalyst bioreactor ; Biocatalyst bioreactor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A simple and efficient method of conversion of wheat starch B to ethanol was investigated. Employing a two-stage enzymatic saccharification process, 95% of the wheat starch was converted to fermentable sugars in 40 h. From 140 g/l total sugars in the feed solution, 63.6 g/l ethanol was produced continuously with a residence time of 3.3 h in a continuous dynamic immobilized biocatalyst bioreactor by immobilized cells ofSaccharomyces cerevisiae. The advantages and the application of this bioreactor to continuous alcoholic fermentation of industrial substrates are presented.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569698
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  • 3
    Electronic Resource
    Electronic Resource
    Manganese accumulation in yeast cells (1989)
    Springer
    BioMetals 2 (1989), S. 6-10 
    Details
    ISSN: 1572-8773
    Keywords: Manganese ; Electron spin resonance ; Superoxide dismutase ; Saccharomyces cerevisiae ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Manganese accumulation was studied by room-temperature electron spin resonance (ESR) spectroscopy inSaccharomyces cerevisiae grown in the presence of increasing amounts of MnSO4. Mn2+ retention was nearly linear in intact cells for fractions related to both low-molecular-mass and macromolecular complexes (‘free’ and ‘bound’ Mn2+, respectively). A deviation from linearity was observed in cell extracts between the control value and 0.1 mM Mn2+, indicating more efficient accumulation at low Mn2+ concentrations. The difference in slopes between the two straight lines describing Mn2+ retention at concentrations lower and higher than 0.1 mM, respectively, was quite large for the free Mn2+ fraction. Furthermore it was unaffected by subsequent dialyses of the extracts, showing stable retention in the form of low-molecular-mass complexes. In contrast, the slope of the line describing retention of ‘bound’ Mn2+ at concentrations higher than 0.1 mM became less steep after subsequent dialyses of the cell extracts. This result indicates that the macromolecule-bound Mn2+ was essentially associated with particulate structures. In contrast to Cu2+, Mn2+ had no effect on the major enzyme activities involved in oxygen metabolism except for a slight increase of cyanide-resistant Mn-superoxide dismutase activity, due to dialyzable Mn2+ complexes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01116194
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  • 4
    Electronic Resource
    Electronic Resource
    Heterologous gene expression of the glyphosate resistance marker and its application in yeast transformation (1989)
    Springer
    Current genetics 15 (1989), S. 91-98 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Vector ; Glyphosate resistance ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The E. coli aroA gene was inserted between yeast promoter and terminator sequences in different shuttle expression plasmids and found to confer enhanced EPSP synthase activity as well as resistance to glyphosate toxicity. Subsequently, a transformation system using these newly constructed vectors in yeast was characterized. The efficiency of the glyphosate resistance marker for transformation and selection with plasmid pHR6/20-1 in S. cerevisiae laboratory strain SHY2 was found to be relatively high when compared with selection for LEU2 prototrophy. The fate of the recombinant plasmid pHR6/20-1 in the transformants, the preservation of the aroA E. coli DNA fragment in yeast, mitotic stability, EPSP synthase activity, and growth on glyphosate-containing medium have been investigated. As this plasmid also allows direct selection for glyphosate resistant transformants on rich media, the glyphosate resistance marker was used for transforming both S. cerevisiae laboratory strain SHY2 and brewer's yeast strains S. cerevisiae var. “uvarum” BHS5 and BHS2. In all cases, the vector pHR6/20-1 was maintained as an autonomously replicating plasmid. The resistance marker is, therefore, suitable for transforming genetically unlabeled S. cerevisiae laboratory, wild, and industrial yeast strains.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00435454
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  • 5
    Electronic Resource
    Electronic Resource
    A hot-spot for transposition of various Ty elements on chromosome V in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 16 (1989), S. 247-252 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Chromosome V ; Ty elements ; tRNA genes ; Transposition hot-spots ; Yeast polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Ty4 is a novel transposable element in the yeast, Saccharomyces cerevisiae, which is present in only a few copies in the genome (Stucka et al. 1989). In strain C836 one of the three copies (Ty4-90) is contained in cosmid clone c90, where it resides on chromosome V. Analysis of this region reveals a “hot-spot” of transposition: in addition to Ty4-90, the locus contains a complete Ty3 element and seven singular delta, sigma and tau elements. Three tRNA genes (for His, Lys, and Ile) are located in this region, and these are closely associated with one or the other of the elements, a phenomenon commonly observed in yeast. A comparison of c90 with corresponding regions from other strains shows that the locus is highly polymorphic and that this polymorphism is explicitly associated with Ty transposition and recombination events.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00422110
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  • 6
    Electronic Resource
    Electronic Resource
    Mutations in ADE3 reduce the efficiency of the omnipotent suppressor sup45-2 (1989)
    Springer
    Current genetics 16 (1989), S. 315-321 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Antisuppressor ; ADE3 ; Translation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutations in a known yeast gene, ADE3, were shown to act as an antisuppressor, reducing the efficiency of the omnipotent suppressor, sup45-2. The ADE3 locus encodes the trifunctional enzyme C1-tetrahydrofolate synthase, which is required for the biosynthesis of purines, thymidylate, methionine, histidine, pantothenic acid and formylmethionyl-tRNAfmet. The role of this enzyme in translational fidelity had not previously been suspected.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340709
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  • 7
    Electronic Resource
    Electronic Resource
    Two new loci that give rise to dominant omnipotent suppressors in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 16 (1989), S. 323-330 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Saccharomyces cerevisiae ; Nonsense suppression ; Omnipotent suppressors ; Gene mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Ten dominant omnipotent suppressors of Saccharomyces cerevisiae, which were previously shown to be different from SUP46, have been examined. Nine are mapped in a region between lys5 and cyh2 on the left arm of chromosome VII. These suppressors, like SUP46, manifest sensitivity to increased temperature and the antibiotics paromomycin and hygromycin B. In addition, they have an identical action spectrum. These results strongly suggest that they are allelic to each other and they are designated SUP138. The tenth is mapped to a position between his1 and arg6 on the right arm of chromosome V. This suppressor, named SUP139, does not manifest temperature sensitivity nor antibiotic sensitivity. SUP139 and SUP138, which are clearly distinguished by means of action spectrum, act on much fewer nonsense mutations than SUP46. It is now clear that dominant omnipotent suppressors arising at a single locus are homogeneous and that their efficiency is locus-dependent. The order of efficiency is SUP46〉SUP138〉SUP139.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340710
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  • 8
    Electronic Resource
    Electronic Resource
    The Saccharomyces cerevisiae RAD2 gene complements a Schizosaccharomyces pombe repair mutation (1989)
    Springer
    Current genetics 15 (1989), S. 27-30 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Repair ; Complementation ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; Gene cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two Saccharomyces cerevisiae genes necessary for excision repair of UV damage in DNA, RAD1 and RAD2, were introduced individually, on a yeast shuttle vector, into seven Schizosaccharomyces pombe mutants — rads1, 2, 5, 13, 15,16 and 17. The presence of the cloned RAD1 gene did not affect survival of any of the S. pombe mutants. The RAD2 gene increased survival of S. pombe rad13 to near the wild-type level after UV irradiation and had no effect on any of the other mutants tested. S. pombe rad13 mutants are somewhat defective in removal of pyrimidine dimers so complementation by the S. cerevisiae RAD2 gene suggests that the genes may code for equivalent proteins in the two yeasts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00445748
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  • 9
    Electronic Resource
    Electronic Resource
    Introduction of nonselectible 2μ plasmid into [cir°] cells of the yeast S. cerevisiae by DNA transformation and in vivo site-specific resolution (1989)
    Springer
    Current genetics 15 (1989), S. 83-90 
    Details
    ISSN: 1432-0983
    Keywords: DNA transformation ; Saccharomyces cerevisiae ; Site-specific recombination ; 2μ DNA plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The 2μ DNA plasmid of the yeast Saccharomyces cerevisiae does not confer any known selectable phenotype to the host cell carrying it. Selection of cells transformed with purified 2μ DNA therefore cannot be achieved, and the intracellular presence of 2μ can only be assessed by molecular analysis of the DNA complement. In addition, 2μ alone does not replicate in bacterial hosts, thus rendering its amplification by conventional methods impossible. We have isolated a shuttle plasmid, pBH-2L, generated by in vivo sites-pecific recombination between the endogenous 2μ DNA plasmid and pRL, a pBR322 derivative containing the yeast LEU2 gene and one 2μ repeat sequence associated with the origin of replication. This new shuttle plasmid has the property, when transformed into yeast, of undergoing site-specific recombinational resolution between its two direct repeat sequences. This releases 2μ plasmid and pRL as individual molecules. The latter can undergo progressive mitotic loss during growth in nonselective medium, ultimately leaving leucine auxotrophic transformants that contain only 2μ DNA plasmid. This system can be utilized to introduce 2μ DNA alone into cells lacking it, thereby providing a novel means to study the biology and the molecular genetics of the plasmid and its potential practical applications as a vector.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00435453
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  • 10
    Electronic Resource
    Electronic Resource
    A galactose-dependent cmd1 mutant of Saccharomyces cerevisiae: involvement of calmodulin in nuclear division (1989)
    Springer
    Current genetics 15 (1989), S. 113-120 
    Details
    ISSN: 1432-0983
    Keywords: Calmodulin mutant ; Nuclear division ; Chromosome stability ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The coding region of a yeast calmodulin gene was fused to a galactose-inducible GAL1 promoter, and a conditional-lethal mutant of Saccharomyces cerevisiae, in which the expression of calmodulin was regulated by galactose, was constructed. The mutant grew normally in galactose medium, but in glucose medium, in which the promoter was repressed, it ceased growing after 12–15 h. The growth arrest was associated with a decrease in intracellular calmodulin levels: after 12h, no intracellular calmodulin protein was detectable. Analysis of the terminal phenotype showed that when the cell stopped growing, it had a bud, a nucleus after S-phase and a short mitotic spindle. Thus, the defect was mainly in nuclear division. Bud growth was partially inhibited in these cells: 27% of the cells stopped growing with a small bud. Furthermore, calmodulin-deficient cells showed elevated rates of chromosome loss, possibly as the result of a defect in the precise segregation of chromosomes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00435457
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  • 11
    Electronic Resource
    Electronic Resource
    A rapid and simple method for extracting yeast mitochondrial DNA (1989)
    Springer
    Current genetics 15 (1989), S. 235-237 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondrial DNA ; Method of extraction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A rapid method for the extraction of yeast mitochondrial DNA (mtDNA) is described. In comparison with previous methods, it simplifies several steps, does not require either the isolation of mitochondria or phenol treatment and is less time consuming. This protocol gives a high yield of pure mtDNA (50–120 μg from a 100-ml culture), which can be directly used in various molecular applications: restriction enzyme digestion, electrophoresis, blotting, labeling, cloning and sequencing.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00435511
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  • 12
    Electronic Resource
    Electronic Resource
    Molecular characterization of the two genes SNQ and SFA that confer Hyperresistance to 4-nitroquinoline-N-oxide and formaldehyde in Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 16 (1989), S. 65-74 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mutagen hyperresistance ; Southern, Northern analysis ; Gene transplacement ; Transposon mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The genes SNQ and SFA confer hyperresistance to 4-NQO and FA when present on a multi-copy plasmid in yeast. Both are non-essential genes since transplacement of SNQ by a disrupted snq-0::LEU2 yielded stable and viable haploid integrants. Southern analysis revealed that SNQ and SFA are single-loci genes, and OFAGE analysis showed that they are located on chromosome XIII and IV, respectively. Northern blot analysis of SNQ and SFA revealed poly(A)+ RNA transcripts of 2 kb and 1.7 kb, respectively. Nuclease S 1 mapping showed SNQ to have a coding region of 1.6 kb and SFA, one of 1.3 kb. The 5′ coding regions were determined for both genes, while the 3′ end could only be determined for gene SNQ. Both genes do not appear to contain introns. The SFA locus was also mapped by transposon mutagenesis. Tn10-LUK integrants disrupted the SFA gene function at sites that were determined by subcloning to lie within the SFA transcription unit.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00393397
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  • 13
    Electronic Resource
    Electronic Resource
    Cloning and characterization of the SKI3 gene of Saccharomyces cerevisiae demonstrates allelism to SKI5 (1989)
    Springer
    Current genetics 16 (1989), S. 139-143 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; SKI3 ; SKI5 ; M1 dsRNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have identified a mutant strain of the yeast Saccharomyces cerevisiae which overproduces killer toxin. This strain contains a single mutation which fails to complement defects in both the SKI3 and SKI5 genes, while a cloned copy of this gene complements both the ski3 and ski5 defects. The level of secreted toxin from a cDNA based plasmid is not increased in a ski3 strain, showing that the overproduction phenotype is dependent upon an increased level of M1 dsRNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00391469
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  • 14
    Electronic Resource
    Electronic Resource
    Reassessing the genotoxic potential of 8-MOP + UVA-induced DNA damage in the yeast Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 16 (1989), S. 75-80 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Psoralen photoaddition ; Interstrand cross-link ; Repair deficiency ; Genotoxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two different UVA irradiation systems were initially biologically calibrated with two haploid yeast strains proficient and deficient, respectively, in nucleotide excision repair. The number of DNA lesions introduced into the cell's genome by the photoactivated bifunctional furocoumarin 8-MOP was then calculated by means of the applied UVA exposure doses. At LD37 the repair-proficient wild type had about 14 ICL and 34 furan-side monoadducts in its DNA, while doubly blocked repair mutant rad3-12 pso1-1 had 2 ICL and 3 monoadducts. Locus-specific reversion of lys1-1 followed two-hit kinetics in the repair-proficient wild type and one-hit kinetics in an excision-deficient rad2-20 mutant, as would be expected if ICL was the main type of mutagenic lesion in the wild type and monoadducts the main mutagenic lesion type in the excision-deficient strain. Quantitative comparison of 8-MOP + UVA-induced ICL with those induced by bifunctional mustard revealed the former to have a much higher genotoxicity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00393398
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  • 15
    Electronic Resource
    Electronic Resource
    The α-factor enhancement of hybrid formation by protoplast fusion in Saccharomyces cerevisiae can be mimicked by other procedures (1989)
    Springer
    Current genetics 15 (1989), S. 303-305 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Protoplast fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The percentage of hybrids formed during protoplast fusion in Saccharomyces cerevisiae is determined by the percentage of protoplasts at the GI/S boundary of the cell cycle.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00447049
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  • 16
    Electronic Resource
    Electronic Resource
    Biosynthesis of the peroxisomal dihydroxyacetone synthase from Hansenula polymorpha in Saccharomyces cerevisiae induces growth but not proliferation of peroxisomes (1989)
    Springer
    Current genetics 16 (1989), S. 13-20 
    Details
    ISSN: 1432-0983
    Keywords: Peroxisomes ; Protein import ; Saccharomyces cerevisiae ; Hansenula polymorpha
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The DAS gene of Hansenula polymorpha was expressed in Saccharomyces cerevisiae under the control of different promoters. The heterologously synthesized dihydroxyacetone synthase (DHAS), a peroxisomal enzyme in H. polymorpha, shows enzymatic activity in baker's yeast. The enzyme was imported into the peroxisomes of S. cerevisiae not only under the appropriate physiological conditions for peroxisome proliferation (oleic acid media), but also in glucose-grown cells where it induced the enlargement of the few peroxisomes present. This growth process was not accompanied by an increase in the number of microbodies, which suggests a separate control mechanism for peroxisomal proliferation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00411078
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  • 17
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    A DNA sequence in Saccharomyces exiguus is homologous with the HO gene of Saccharomyces cerevisiae (1989)
    Springer
    Current genetics 15 (1989), S. 399-401 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces exiguus ; Saccharomyces cerevisiae ; HO gene ; MAT gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The DNA of Saccharomyces exiguus was analyzed by Southern hybridization using cloned MATa, MATα, and HO genes of Saccharomyces cerevisiae as probes. It was shown that S. exiguus has a DNA sequence homologous with the HO gene of S. cerevisiae and that this DNA sequence is on a chromosome of about 940 kb of DNA in S. exiguus. However, there is no DNA sequence in S. exiguus that is homologous with the MAT genes of S. cerevisiae.
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    URL: http://dx.doi.org/10.1007/BF00376794
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  • 18
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    Kinetic analysis and simulation of glucose transport in plasma membrane vesicles of glucose-repressed and derepressedSaccharomyces cerevisiae cells (1989)
    Springer
    Cellular and molecular life sciences 45 (1989), S. 1018-1023 
    Details
    ISSN: 1420-9071
    Keywords: Saccharomyces cerevisiae ; growth conditions ; kinaseless mutant ; plasma membrane vesicles ; glucose transport ; kinetics and computer simulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In this study experimental data on the kinetic parameters investigated by other authors1–5, 11 together with own data on plasma membrane vesicles, have been subjected to a computer simulation based on the equations describing facilitated diffusion. The simulation led to an ideal fit describing the above data. From this it can be concluded that glucose is transported by facilitated diffusion, and not by active transport as was postulated by Van Steveninck14, 15. The simulation method also demonstrates that the fast sampling technique used by these authors1–5,11 underestimates the fluxes. Thus, the parameters given do not contribute to the understanding of glucose transport under different metabolic conditions. The K value of plasma membrane vesicles prepared from glucose-repressed cells is around 7 mM. Derepression, particularly by galactose, causes a highly significant increase in affinity as shown by a decrease in the K value to 2 mM. The highest affinity was measured in a triple kinaseless mutant grown on glycerol with a K value of 1 mM. If seems, therefore, that the kinetic parameters derived from initial uptake rates of glucose in intact cells1–5,11 using single flux analysis, such as Eadie-Hofstee- or Lineweaver-Burk-plots, are in error.
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    URL: http://dx.doi.org/10.1007/BF01950152
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  • 19
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    Effects of nucleotides and divalent cations on phospholipase activity in Saccharomyces cerevisiae (1989)
    Springer
    Archives of microbiology 151 (1989), S. 154-158 
    Details
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Yeast ; Phospholipase B ; Lysophospholipase ; Enzyme inhibition ; AMP ; Unesterified fatty acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Divalent cations activate the lysophospholipase and transacylase reactions catalyzed by the same enzymes in the yeast Saccharomyces cerevisiae. The activation was observed at neutral pH, but not at the pH optimum of lysophospholipase/transacylase, near 3.5. Adenine nucleotides, especially AMP and ADP, are strong inhibitors of the same group of enzymes. Half maximal inhibition by AMP was found at a concentration of about 20 μM. The inhibition by nucleotides in low concentrations is enhanced by divalent cations.
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    URL: http://dx.doi.org/10.1007/BF00414431
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  • 20
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    Genetic analysis of inducible sexual agglutination ability in the yeast Saccharomyces cerevisiae (1989)
    Springer
    Archives of microbiology 151 (1989), S. 198-202 
    Details
    ISSN: 1432-072X
    Keywords: Sexual agglutination ; Mating ; Saccharomyces cerevisiae ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic regulation of the inducibility of sexual agglutination ability in the yeast Saccharomyces cerevisiae was studied. Detailed analysis of the degree of sexual agglutination was carried out; it showed that a greater number of genes are involved in the regulation of inducible sexual agglutination in strain H1-0 than previously assumed. Although dominancy of inducible phenotype over constitutive was confirmed, the effectiveness of one gene changing the constitutive phenotype to the inducible seemed to be somewhat low. Quantity per cell of agglutination substances responsible for sexual agglutination increased as the agglutination ability became greater.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00413130
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  • 21
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    A similar protein portion for two exoglucanases secreted by Saccharomyces cerevisiae (1989)
    Springer
    Archives of microbiology 151 (1989), S. 391-398 
    Details
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Exoglucanases ; Purification ; Protein moieties ; Tunicamycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Exoglucanase (exo-1,3-β-D-glucan glycohydrolase, EC 3.2.1.56) activity secreted by Saccharomyces cerevisiae into the culture medium was separated by ion exchange chromatography into two glycoprotein isoenzymes which contributed 10% (exoglucanase I) and 90% (exoglucanase II) towards the total activity. Analysis of the “in vitro” deglycosylated products by polyacrylamide gel electrophoresis under native or denaturing conditions indicated that the protein portions of both exoglucanases exhibited identical mobility, each one consisting of two polypeptides with M r of 47000 and 48000. The same profile was shown by the exoglucanase secreted in the presence of tunicamycin. Antibodies raised against the protein portion of exoglucanase II did react with both native exoglucanases and their deglycosylated products with a pattern indicative of immunological identity. Digestion of the “in vitro” deglycosylated products of both exoglucanases with Staphylococcus aureus V-8 protease or trypsin generated the same proteolytic fragments in each case. Only exoglucanase II was secreted by protoplasts. These and previously reported results indicate that the protein portions of both isoenzymes may be the product of the same gene (or a family of related genes), and that exoglucanase I is a product of enzyme II, modified by a process occurring beyond the permeability barrier of the cell.
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    URL: http://dx.doi.org/10.1007/BF00416596
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  • 22
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    Cloning of the LEU2 gene of Saccharomyces cerevisiae by in vivo recombination (1989)
    Springer
    Archives of microbiology 152 (1989), S. 263-268 
    Details
    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Recombination ; Tryptophan cluster ; Yeast vectors ; Plasmid copy number
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We describe a convenient method for the in vivo construction of large plasmids that possess a multitude of restriction sites. A large (23 kbases) circular self-replicating plasmid carrying a partial LEU2-d gene was cotransformed with a circular non-replicating plasmid carrying the entire LEU2 gene. In vivo recombination results preferentially in a plasmid that carries both the LEU2-d and the entire LEU2 gene. In addition we also found one plasmid with a tandem LEU2 insertion and one plasmid where the LEU2-d gene was replaced by the entire LEU2 gene.
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    URL: http://dx.doi.org/10.1007/BF00409661
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    GABA-like immunoreactivity in the suboesophageal ganglion of the locust Schistocerca gregaria (1989)
    Springer
    Cell & tissue research 258 (1989), S. 53-63 
    Details
    ISSN: 1432-0878
    Keywords: GABA ; Immunohistochemistry ; Salivary neurones ; Schistocerca gregaria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neurones in the suboesophageal ganglion of the locust Schistocerca gregaria were stained with an antiserum raised against gamma amino butyric acid (GABA). This ganglion consists of the fused mandibular, maxillary and labial neuromeres. Immunoreactive cell bodies of similar size and distribution occur in the lateral, ventral and middorsal regions of all three neuromeres. Approximately 200 cell bodies stain in both the mandibular and maxillary neuromeres and 270 in the labial neuromere. A few distinctly larger cells occur in the ventral groups and one large pair occurs in the lateral group of the maxillary neuromere. Dorsal commissures DCIV and DCV are composed mainly of stained fibres, while DCI–DCIII are largely unstained. A ventral commissure also stains in the maxillary neuromere. All longitudinal tracts contain both stained and unstained fibres. Many processes within the neuropil are also immunoreactive. A stained axon is found in the posterior tritocerebral commissure which enters the anterior dorsal region of the mandibular neuromere. The salivary branch of the 7th nerve contains one stained axon and two axons stain in nerve 8 which innervates neck muscles.
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    URL: http://dx.doi.org/10.1007/BF00223144
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    The distribution of endocrine cells in the mucosa of the gastrointestinal tract of the house musk shrew, Suncus murinus (Insectivora) (1989)
    Springer
    Cell & tissue research 258 (1989), S. 365-371 
    Details
    ISSN: 1432-0878
    Keywords: Gastrointestinal tract ; Gastric mucosa ; Gastrointestinal endocrine cells ; Immunohistochemistry ; Suncus murinus (Insectivora)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of endocrine cells in the gastrointestinal tract of the house musk shrew, Suncus murinus (Family Soricidae, Order Insectivora) was studied immunohistochemically. The hormones investigated were gastrin, cholecystokinin (CCK), somatostatin, secretin, glucagon, gastric inhibitory polypeptide (GIP), motilin and neurotensin. In the gastric mucosa, gastrin and somatostatin cells were only found in the pyloric regions, and no other hormonal cell-types were observed. In the intestinal mucosa, the largest number of endocrine cells belonged to the gastrin and glucagon/glicentin cell-types, whereas CCK-33/39 and secretin cells were the least numerous. Numbers of other cell-types were intermediate between these two groups. The gastrin and GIP cells were mostly localized in the proximal portion of the intestine, decreasing in number towards the distal portion. The motilin and CCK-33/39 cells were restricted to the proximal half. The glucagon/glicentin and neurotensin cells were most abundant in the middle portion. The somatostatin and secretin cells, although only present in small numbers, were randomly distributed throughout the intestine. This characteristic distribution of gastrointestinal endocrine cells is discussed in comparison with the distribution patterns of other mammals.
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    URL: http://dx.doi.org/10.1007/BF00239456
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    The blood-testis barrier and its breakdown following immunization to testis material in the Nile tilapia, Oreochromis niloticus (1989)
    Springer
    Cell & tissue research 258 (1989), S. 491-498 
    Details
    ISSN: 1432-0878
    Keywords: Blood-testis barrier ; Immune responses ; Tracer studies ; Immunohistochemistry ; Oreochromis niloticus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The blood-testis barrier and its changes following immunization to testis material, were investigated by light- and electron microscopy in a teleost fish, the Nile tilapia Oreochromis niloticus, using horseradish peroxidase and bovine serum albumin as tracers. In the normal testis, histochemistry using horseradish peroxidase revealed that a barrier composed of junctional complexes connecting adjacent Sertoli cells existed around the central lumina of the seminal lobules, and also around the germ-cell cysts containing spermatids at the middle or late phase of chromatin condensation. By contrast, bovine serum albumin was prevented from passing through the basement membrane and could not penetrate any of the spermatogenetic cysts, indicating that the basement membrane may be an ion-selective barrier. In tilapia immunized with allogeneic testis homogenate emulsified in Freund's complete adjuvant, bovine serum albumin could penetrate the spermatogenetic cysts, and horseradish peroxidase was able to pass through the intercellular spaces between Sertoli cells to the region nearer the seminal lobule lumen, due to the junctional complexes becoming loosened. The results suggest that the blood-testis barrier, both junctional complexes and the basement membrane, are broken down during immune responses.
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    URL: http://dx.doi.org/10.1007/BF00218861
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    Ectopic Purkinje-like cells are GABAergic: Immunohistochemistry with an immune serum against glutamic acid decarboxylase (1989)
    Springer
    Cell & tissue research 258 (1989), S. 437-439 
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    ISSN: 1432-0878
    Keywords: Cerebellum ; Purkinje cells ; Ectopia ; GABA ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Intensely stained cells are found in the cerebellar white matter of the vermis and paravermis in adult rats after immunoreaction with an immune serum raised against glutamic acid decarboxylase (GAD). The cells are similar in size to cortical Purkinje cells and three times the size of Golgi cells of the internal granule layer, and have a thick immunopositive cell process emerging from a welldefined cytoplasmic cone. In the cytoplasm, immunoprecipitates are more dense around the nucleus as in normally located Purkinje cells. The morphological appearance of the immunopositive cells suggests that they may be ectopically located Purkinje cells. The soma of the ectopic Purkinje cells is contacted by a few darkly stained terminal boutons. Data indicate that, in spite of the different cellular environment, ectopic Purkinje cells can develop not only the typical morphological pattern already described but also other intrinsic features, such as their typical inhibitory neurotransmitter.
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    URL: http://dx.doi.org/10.1007/BF00239466
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    Localization of corticotropin-releasing hormone (CRF)-like immunoreactivity in the central nervous system of the elasmobranch fish, Scyliorhinus canicula (1989)
    Springer
    Cell & tissue research 258 (1989), S. 541-546 
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    ISSN: 1432-0878
    Keywords: Corticotropin-releasing factor hormone ; Immunohistochemistry ; Radioimmunoassay ; Hypothalamus ; Scyliorhinus canicula (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The occurrence and localization of immunoreactive corticotropin-releasing factor (CRF) in the brain and pituitary of the elasmobranch fish Scyliorhinus canicula, were studied by means of specific radioimmunoassay and immunohistochemistry using the indirect immunofluorescence method. Brain and pituitary extracts showed a good cross-reactivity with the ovine CRF antiserum, but serial dilutions of tissue samples did not completely parallel the standard curve. Relatively high concentrations of CRF-like material were found within the pituitary, diencephalon, and telencephalon. CRF-like immunoreactive perikarya were observed in the preoptic nucleus and in the nucleus lateralis tuberis. Numerous immunoreactive cells appeared to be of the CSF-contacting type. CRF-like immunopositive fibers were seen to run through the hypothalamus within the ventro-medial floor of the infundibular region. A dense plexus of immunoreactive nerve endings terminated in the median eminence and the neurointermediate lobe of the pituitary. These results indicate that a neurosecretory system containing CRF-like immunoreactivity exists in the brain of elasmobranchs, a group of vertebrates which has diverged early from the evolutionary line leading to mammals. In addition, our data support the notion that a CRF-like molecule is involved in the regulation of corticotropic and melanotropic cell activity in this primitive species of fish.
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    URL: http://dx.doi.org/10.1007/BF00218865
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    An immunohistochemical study on the postnatal development of rat nasal-associated lymphoid tissue (NALT) (1989)
    Springer
    Cell & tissue research 256 (1989), S. 431-438 
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    ISSN: 1432-0878
    Keywords: Mucosa ; Lymphoid tissue ; Nose ; Development ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This study concerns the development of nasal-associated lymphoid tissue in the rat, using immuno- and enzyme-histochemical staining techniques on cryostat sections. Nasal-associated lymphoid tissue is present at birth as a small accumulation of mainly T lymphocytes and non-lymphoid cells; B cells are rare. Distinct areas of T and B cells appear at 10 days after birth; by that time high endothelial venules are also observed. Intra-epithelial lymphocytes are present, most of them being T-helper cells. ED1+ macrophages are seen throughout the tissue. The proportion of ED1+cells does not change during ontogeny. ED2+cells (tissue macrophages) are present predominantly at the border between the lymphoid tissue and the surrounding connective tissue, in all age-groups. ED3+mononuclear cells are scattered throughout the nasal-associated lymphoid tissue of young animals. Later on, the ED3+ cells migrate into the border-area between lymphoid and connective tissue. Ia+ non-lymphoid cells in the nasal lymphoid tissue increase in number during ontogeny. Only a few of them show acid phosphatase activity, indicating that the proportion of classical scavenger macrophages is low. Some of them may be antigen presenting (dendritic) cells. Ia+ dendritic cells also occur between the epithelial cells. Moreover, some epithelial cells express the Ia marker.
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    URL: http://dx.doi.org/10.1007/BF00218901
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    Distribution of FMRFamide-like immunoreactivity in the brain and neurohypophysis of the lamprey, Lampetra japonica (1989)
    Springer
    Cell & tissue research 256 (1989), S. 581-584 
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    ISSN: 1432-0878
    Keywords: FMRFamide ; Brain ; Neurohypophysis ; Pineal body ; Immunohistochemistry ; Lampetra japonica (Cyclostomata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Distribution of molluscan cardio-excitatory tetrapeptide Phe—Met—Arg—Phe—NH2 (FMRFamide) was determined by means of immunohistochemistry in the brain and neurohypophysis of the lamprey, Lampetra japonica. Many FMRFamide-like immunoreactive neurons were found in the periventricular nuclear region and in a region near the mammillary recess. Neurons situated in the former region were larger. The immunoreactive cell groups were shown to be located at sites differing from those of the AF-positive cell groups. The fibers of immunoreactive neurons extended in all directions within the brain and towards the spinal cord, some reaching the third ventricle and capillaries. Thus, FMRFamide-like immunoreactive peptides appear to function as neurotransmitters or neuromodulators and possibly also as neurohormones. FMRFamide-like immunoreactive material was rarely observed in the posterior neurohypophysis (neural lobe), but was noted to be present to a limited extent in the caudal part of the anterior neurohypophysis (median eminence). It would thus follow that FMRFamide-like immunoreactive neurons may not necessarily be related to the hypothalamo-neural lobe system, but may possibly be associated with the hypothalamoadenohypophysial system. The pineal body showed no FMRFamide-like immunoreactivity.
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    URL: http://dx.doi.org/10.1007/BF00225607
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    Origin of the met-enkephalinergic innervation of the lateral septum in the rat (1989)
    Springer
    Cell & tissue research 256 (1989), S. 585-592 
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    ISSN: 1432-0878
    Keywords: Axonal retrograde tracing ; Hypothalamus ; Immunohistochemistry ; Methionine-enkephalin ; Septum ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The location of the cells giving rise to the methionine-enkephalin (Met-Enk)-ergic innervation of the lateral septal nucleus has been investigated in the rat by combining immunohistochemistry and retrograde axonal tracing. Small volumes (0.06 μl) of apo-horseradish peroxidase (Apo-HRP) conjugated to wheat-germ agglutinin (WGA) and coupled with colloidal gold particles (WGA-ApoHRP-gold) were injected into the lateral septum. The retrogradely labeled cell bodies were visualized by silver intensification of the gold particles on Vibratome sections that were subsequently processed for immunohistochemistry for Met-Enk. Cells labeled with WGA-ApoHRP-gold were observed in the septal area, throughout the hypothalamus (mainly in the perifornical and lateral nuclei) and in the mesencephalon. The localization of Met-Enk-immunoreactive cells was as previously described. With the exception of a few septal cells close to the injection site, doubly labeled cells were found only in the perifornical nucleus of the hypothalamus. Almost all perifornical magnocellular cells were doubly labeled ipsilateral to the injection site, whereas on the opposite side, only about 25% of the Met-Enk-immunoreactive cells contained WGA-ApoHRP-gold. Other brain regions containing retrogradely labeled or Met-Enk-immunoreactive cells (particularly the raphe nuclei) did not show double-labeled neurons. This study demonstrates, using a new and sensitive technique for specific neurochemical tracing of tracts, that the origin of the Met-Enk-ergic innervation of the rat lateral septal nuclei lies in the magnocellular perifornical nuclei of the hypothalamus. The precise involvement of this pathway in limbic functions remains to be determined.
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    URL: http://dx.doi.org/10.1007/BF00225608
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    Characterization of the follicles in the intermediate lobe of the pituitary gland of the Mongolian gerbil (Meriones unguiculatus) (1989)
    Springer
    Cell & tissue research 257 (1989), S. 657-660 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary ; Intermediate lobe follicles ; Immunohistochemistry ; α-MSH ; Agranular cells ; Meriones unguiculatus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The Mongolian gerbil (Meriones unguiculatus) contains abundant follicles throughout the intermediate lobe (IL) of the pituitary gland in the adult animal. The mode of follicle formation, the nature of the follicle building cells and the distribution of follicles were investigated in semithin sections of the gerbil IL. The sections were stained conventionally, or immunohistochemically with antibodies directed against α-melanocyte stimulating hormone (α- MSH). Follicular cells were constantly α-MSH-negative, and resembled the marginal cells lining the hypophyseal cleft with regard to their cytological and immunohistochemical properties. Moreover, follicular cells appeared to be derived from strands of marginal cells that regularly invaginated deep into the IL. Both follicular and marginal cells often made up cellular clusters. This process coincided with follicle formation and the generation or transport of the colloidal content found inside follicles and the hypophyseal cleft. Although the non-secretory cells of the IL obviously constituted one major source of pituitary colloid in the gerbil, α-MSH-positive secretory cells, which occasionally were found to be discharged into the cleft cavity, might contribute to the colloidal contents.
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    URL: http://dx.doi.org/10.1007/BF00221478
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    Gastrin/cholecystokinin-like immunoreactivity in the nervous system of Aeschna cyanea (Insecta, Odonata) (1989)
    Springer
    Cell & tissue research 257 (1989), S. 105-113 
    Details
    ISSN: 1432-0878
    Keywords: Insect nervous system ; Cholecystokinin-like peptide ; Immunohistochemistry ; Radioimmunoassay ; Aeschna cyanea (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Gastrin/cholecystokinin (gastrin/CCK)-like immunoreactivity has been detected in the brain, suboesophageal ganglion and corpora cardiaca of the larva of Aeschna cyanea by radioimmunoassay and immunohistochemistry, by use of two antisera raised against the sulfated (CCK-8S) and the unsulfated form (CCK-8NS) of the carboxyl terminal octapeptide. Numerous immunoreactive neurons were demonstrated in the protocerebrum (exclusive of optic lobes) and suboesophageal ganglion where 20 and 15 symmetrical clusters of reactive cells, respectively, were observed. Immunoreactive cells also occurred in the tritocerebrum, the optic lobes and the frontal ganglion. In the corpora cardiaca, gastrin/CCK-like material was found both within intrinsic cells and axon terminals. RIA measurements support the immunohistochemical results in so far as large amounts of gastrin/CCK-like material were detected in the brain, corpora cardiaca and suboesophageal ganglion complex. Both boiling water-acetic acid- and methanol-extraction procedures were performed. Comparisons of the results lead to the conclusion that a large part of the gastrin/CCK-like material occurs as small molecules. Immunohistochemical procedures performed on material fixed in a solution of picric acid-paraformaldehyde demonstrated differences in the immunoreactivity of the tested antisera. First, the immunohistochemical reaction was always more pronounced when the CCK-8NS antiserum was used instead of the CCK-8S antiserum, which may be interpreted by a lower affinity of the latter. In the second place, some neurons strongly stained by the CCK-8NS antiserum were only very faintly if at all stained by the CCK-8S antiserum, which may mean that different peptides or at least distinct forms of the same precursor are detected.
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    Immunohistochemical study on fetal raphe samples transplanted into the leptomeningeal tissue of 5,6-dihydroxytryptamine-treated adult rats (1989)
    Springer
    Cell & tissue research 256 (1989), S. 457-463 
    Details
    ISSN: 1432-0878
    Keywords: Transplantation ; Serotonin ; Tyrosine hydroxylase ; Immunohistochemistry ; Leptomeninges ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pieces of fetal midbrain raphe containing serotonergic and dopaminergic neurons were transplanted into the leptomeningeal tissue (see Fig. 3) of adult host rats that had previously been denervated by treatment with 5,6-dihydroxytryptamine. One, 2 and 5 months after transplantation, the rate of neuronal survival in the grafted tissue and the extent of axonal outgrowth into the host brain were studied by use of serotonin and tyrosine hydroxylase (TH) immunohistochemistry. The survival rate of the grafts in the 1-month group was approximately 70%. Neurons containing either serotonin or catecholamine were demonstrated by means of immunocytochemical procedures in the grafts. Two and 5 months after transplantation, serotonin-immunoreactive nerve fibers were densely distributed throughout the graft tissue, while TH-immunoreactive fiber elements were restricted to an area near the somata of TH-positive neurons. Numerous serotonin-immunoreactive fibers derived from the transplant were found in the leptomeningeal tissue surrounding the graft, on the wall of neighboring blood vessels, and also in the adjacent parenchyma of the host brain. Outgrowing TH-immunoreactive nerve fibers were not observed in the host brain, although such elements occurred in the leptomeningeal tissue and the wall of the larger blood vessels. These results suggest that the serotonergic and catecholaminergic (dopaminergic) neurons located in transplants of the raphe nuclei show different patterns when reinnervating the host tissue.
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    Two types of mammosomatotropes in mouse adenohypophysis (1989)
    Springer
    Cell & tissue research 256 (1989), S. 645-648 
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    ISSN: 1432-0878
    Keywords: Mammosomatotropes ; Adenohypophysis ; Electron microscopy ; Immunohistochemistry ; Mouse (SMA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two types of mammosomatotropes (MS), the small-granule and vesicle-granule MS, were detected in mouse adenohypophysis by electron microscopy and immunohistochemistry. Both cell-types were immunoreactive to prolactin (PRL) and growth hormone (GH) antisera. The small-granule MS contained small, round, solid secretory granules about 100 nm in diameter, and were smaller than the classical GH and PRL cell-types. The vesicle-granule MS contained secretory granules like cored vesicles, and were larger than classical GH and PRL cells. Small-granule MS were immunoreactive to both PRL and GH antisera in the same region of the cell cytoplasm; the vesicle-granule MS, however, were immunoreactive to only PRL antiserum in most cytoplasmic areas, and a positive response to both PRL and GH antisera was confined to only certain small areas.
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    Smooth muscle fibers of Podocoryne carnea (Hydrozoa) demonstrated by a specific monoclonal antibody and their association with neurons showing FMRFamide-like immunoreactivity (1989)
    Springer
    Cell & tissue research 255 (1989), S. 275-282 
    Details
    ISSN: 1432-0878
    Keywords: Monoclonal antibody ; FMRFamide ; Immunohistochemistry ; Smooth muscle ; Nervous system ; Podocoryne carnea (Cridaria)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A mouse monoclonal antibody was prepared by using homogenized fragments of crude umbrella material of the hydromedusa Podocoryne carnea as an antigen. The selected clone produced an IgG (mAb sm-1) which decorated smooth muscle cells of hydrozoans. Immunohistochemical testing of mAb sm-1 on whole-mount preparations revealed reactivity with a cytoplasmic, formaldehyde-resistant antigen present in the smooth muscle cells, but absent in all other cell-types. The antibody can therefore be used as a selective and highly sensitive marker to trace the pattern of the smooth muscle system in hydrozoans. The tight association between smooth muscle cells and nerve cells which show FMRFamide-like immunoreactivity can be demonstrated in whole-mount preparations of the hydromedusa Podocoryne carnea with a polyclonal anti-FMRFamide antiserum and in double-labelling experiments.
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    URL: http://dx.doi.org/10.1007/BF00224109
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    Phenotypic heterogeneity of vascular endothelial cells in the human kidney (1989)
    Springer
    Cell & tissue research 256 (1989), S. 27-34 
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    ISSN: 1432-0878
    Keywords: Kidney ; Endothelium ; Monocyte ; Von Willebrand factor ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To clarify the structural base of immune response occurring in the kidney, we investigated the antigenic and functional properties of vascular endothelial cells. Peritubular capillary endothelial cells exhibited the same immuno-histochemical characteristics (OKM5-positive, HLA-DR-positive, Factor VIII/von Willebrand factor antigen-negative, Interleukin 1-positive) as a peripheral blood macrophage subset capable of presenting soluble antigens and triggering the autologous mixed lymphocyte reaction. On the other hand, endothelial cells of glomerular capillary loops, considered to be involved in blood coagulation, were OKM5-negative, HLA-DR-positive, Factor VIII/von Willebrand factor antigen-positive, Interleukin 1-positive. Thus the results of this study suggest that vascular endothelial cells in different anatomic compartments of the kidney express surface antigens heterogenously and may play different roles in the immune reaction.
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    Innervation of the gastrointestinal canal of the toad Bufo marinus by neurons containing 5-hydroxytryptamine-like immunoreactivity (1989)
    Springer
    Cell & tissue research 255 (1989), S. 601-609 
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    ISSN: 1432-0878
    Keywords: Gut ; Immunohistochemistry ; Neurons ; Serotonin ; Bufo marinus (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The gut of the toad, Bufo marinus, was examined for evidence of enteric neurons containing 5-hydroxytryptamine-like immunoreactivity. Such neurons were absent from the stomach. They were present in the small intestine, with processes confined to the myenteric plexus. Immunoreactive nerve cell bodies lay on branches of the pelvic nerves supplying the large intestine; fibres were found in the submucosa of the posterior large intestine and in the muscularis externa of the anterior large intestine. It is concluded, on morphological grounds, that the neurons in the small intestine are interneurons, whereas those in the large intestine are postganglionic parasympathetic motoneurons.
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    Isolation of multicellular complexes of follicular dendritic cells and lymphocytes: Immunophenotypical characterization, electron microscopy and culture studies (1989)
    Springer
    Cell & tissue research 257 (1989), S. 9-15 
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    ISSN: 1432-0878
    Keywords: Follicular dendritic cells ; B-lymphocytes ; Follicular structure ; Immunohistochemistry ; Cell culture ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphological and phenotypical features of multicellular complexes formed by follicular dendritic cells and lymphocytes (FDC-LC) isolated from human hyperplastic tonsils and adenoids are described. FDC-LC obtained with this procedure were morphologically and immuno-phenotypically heterogeneous. In one type of FDCLC, probably obtained from germinal centers, the lymphocytes exhibited ultrastructural features of centroblasts and centrocytes. In a second type, likely derived from follicular mantles, the enclosed lymphocytes were small in size and characterized by a condensed chromatin pattern. Similar heterogeneity was observed by immuno-phenotypical analysis, which revealed a prevalence of IgD+, CD3-, MT2+ small lymphocytes in a high proportion of FDC-LC. Both types of FDC-LC contained desmoplakin immunoreactivity in a typical punctate pattern corresponding to intercellular junctions when tested with a specific antibody. These findings confirm the importance of FDC in maintaining the follicular structure and also suggest that the different zones forming lymphoid follicles (mantle zone and germinal center) are formed by lymphocytes gathered in single “domains” by cytoplasmic processes of FDC. These domains have strong resistance to mechanical stress, such as that used in isolation procedures. FDC-LC have also been maintained as organized multicellular clusters for short periods (more than 48 h) in agarose gel cultures.
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    URL: http://dx.doi.org/10.1007/BF00221628
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    A comparative study of neuropeptides in the intestine of two stomachless teleosts (Poecilia reticulata, Leuciscus idus melanotus) under conditions of feeding and starvation (1989)
    Springer
    Cell & tissue research 255 (1989), S. 245-254 
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    ISSN: 1432-0878
    Keywords: Enteroendocrine cells ; Immunohistochemistry ; Bombesin ; Enkephalin ; Gastrin/CCK ; 5-HT ; Neuropeptide Y ; Neurotensin ; Substance P ; VIP ; Intestine ; Starvation ; Leuciscus idus melanotus ; Poecilia reticulata (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Endocrine cells containing bombesin-, enkephalin-, gastrin/CCK-, 5-HT-, and substance P-like material were demonstrated in the alimentary tract of Poecilia reticulata and Leuciscus idus melanotus. Endocrine cells with neuropeptide-Y-like immunoreactivity were found only in P. reticulata, those with VIP-like immunoreactivity only in L. idus melanotus. Gut nerves showing bombesin-, G/CCK-5-HT-, neurotensin-, substance P-and VIP-like immunoreactivity were observed in both species investigated, enkephalin- and neuropeptide Y-like immunoreactivity in P. reticulata alone. The distribution and amount of endocrine cells and nerves along the gut as visualized with the appropriate antisera varied in both teleosts. Histologically, the intestinal tract of these stomachless fish can be divided into three regions. A large number of endocrine cells with VIP-like immunoreactivity was noted in the rectum of L. idus melanotus. Endocrine cells containing bombesin-, enkepha-lin- and substance P-like material were found only in intestinal parts I and II in L. idus melanotus. Neuropeptide Y-like immunoreactivity was absent from intestinal part I of P. reticulata. The influence of starvation on the immunoreactivity of nerves and enteroendocrine cells in the teleost intestine was examined. After a starvation period of more than 6 weeks, no alterations were observed either in the appearance or amount of nerve and endocrine cell immunoreactivity.
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    URL: http://dx.doi.org/10.1007/BF00229087
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    Neuropeptide Y-like immunoreactivity in the cardiovascular nerve plexus of the elasmobranchs Raja erinacea and Raja radiata (1989)
    Springer
    Cell & tissue research 255 (1989), S. 481-486 
    Details
    ISSN: 1432-0878
    Keywords: Neuropeptide Y (NPY) ; Catecholamines ; Cardiovascular system, innervation ; Immunohistochemistry ; Elasmobranchs (Raja erinacea, Raja radiata, Squalus acanthias) ; Teleosts (Gadus morhua, Salmo gairdneri)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of nerves showing neuropeptide Y (NPY)-like immunoreactivity in the cardiovascular system of elasmobranchs and teleosts has been investigated. Two species of teleosts, the rainbow trout (Salmo gairdneri) and the Atlantic cod (Gadus morhua), and three species of elasmobranchs, the spiny dogfish (Squalus acanthias), the little skate (Raja erinacea) and the starry ray (Raja radiata), were used in this study. An innervation of the cardiovascular system by an NPY-like substance was found only in the two species of Raja. A rich innervation was encountered in these skates, with the highest density of fibres in the wall of the ventricle, the conus arteriosus, the coeliac artery and smaller mesenterial vessels. In the vessels, the fibres formed a plexus at the adventitio-mediol border. Few fibres were found in the walls of the dorsal aorta, the sinus venosus and the atrium, and no fibres were observed in the walls of the ventral aorta. Falck-Hillarp fluorescence histochemistry showed the presence of a rich innervation of arteries and arterioles of the gut by catecholamine-containing nerve fibres.
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    URL: http://dx.doi.org/10.1007/BF00218782
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    Immunohistochemical localization of glial fibrillary acidic protein (GFAP) and vimentin in the subcommissural organ of the Mongolian gerbil (Meriones unguiculatus) (1989)
    Springer
    Cell & tissue research 255 (1989), S. 595-600 
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    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Intermediate filaments ; Glial fibrillary acidic protein (GFAP) ; Vimentin ; Immunohistochemistry ; Meriones unguiculatus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The chemical composition of intermediate filaments (IF's) in the ependyma of the subcommissural organ (SCO) of the Mongolian gerbil (Meriones unguiculatus) was investigated immunohistochemically in paraffin-embedded tissue. Antibodies against glial fibrillary acidic protein (GFAP), vimentin, neurofilament proteins and cytokeratins were used. Only GFAP and vimentin were detected in the non-specialized diencephalic ependyma and in the ependymocytes of the SCO. Staining could be observed in apical and basal processes of the SCO-cells. The latter processes extended into the posterior commissure up to the subpial surface, thus establishing a well-developed leptomeningeal route of ependymal projections. In contrast to the homogeneous vimentin-labeling, the SCO was particularly immunoreactive for GFAP in its lateral aspects and in the supraand precommissural parts. The coexpression of GFAP and vimentin in a subclass of SCO-ependymocytes was demonstrated on differentially immunostained semithin sections. The present study confirms the glial nature of the SCO-ependyma, which has been a matter of debate recently. It appears from this investigation that the high degree of secretory activity in the SCO does not necessarily lead to the disappearance of glial IF proteins. Moreover, the SCO-cells belong to the expanding group of mature astroglia, which is characterized by coexpression of GFAP and vimentin. The morphological similarity between SCO-ependymocytes and tanycytes is underscored by their common immunoreactivity against these two IF proteins. In view of the absence of GFAP from the rat SCO, interspecific differences must be considered in the evaluation of the IF protein composition.
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    URL: http://dx.doi.org/10.1007/BF00218796
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    Immunohistochemical localization of tissue-type plasminogen activator in ovaries before and after induced and spontaneous ovulation in the rat (1989)
    Springer
    Cell & tissue research 257 (1989), S. 1-8 
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    ISSN: 1432-0878
    Keywords: Ovary ; Immunohistochemistry ; Plasminogen activator ; Ovulation ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The observation that tissue-type plasminogen activator (tPA) activity increased dramatically in preovulatory follicles has led to the hypothesis that plasminogen activation is causally related to follicle rupture. With immunohistochemistry, we have studied the appearance of tPA in ovaries of immature rats induced to ovulate and in adult cycling rats. Treatment of immature female rats with a single dose of pregnant mare serum gonadotropin (PMSG) induced follicular maturation. A subsequent human chorionic gonadotropin (hCG) injection resulted in follicle rupture 12–14 h later. PMSG treatment alone did not induce appearance of tPA-immunoreactive cells in any ovarian compartment. After hCG stimulation, however, theca cells, granulosa cells, and oocytes of pre- and postovulatory follicles displayed distinct tPA immunoreactivity. Fibroblastlike cells in the theca layers and tunica albuginea of the follicle apex also demonstrated localized cytoplasmic tPA reactivity. In addition to tPA synthesis in preovulatory follicles, hCG also induced tPA staining in the theca (but not granulosa) layers of non-ovulatory follicles. At 24 h after hCG treatment, there was a marked tPA staining in developing corpora lutea, ovulated ova, and oviductal epithelium. Ovaries from regularly cycling adult rats displayed a similar ovulation-related pattern of tPA immunostaining. The appearance of tPA in different cell types of the preovulatory follicle and in the fibroblast-like cells at the follicle apex, strengthens the hypothesis of a direct involvement of tPA in follicle rupture. Presence of tPA in postovulatory oocytes, cumulus cells, and surrounding oviductal epithelium may also indicate a role for tPA in the transfer of eggs in the oviduct.
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    URL: http://dx.doi.org/10.1007/BF00221627
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    Immunohistochemical demonstration and radioimmunoassay of melatonin in the mink pineal gland (1989)
    Springer
    Cell & tissue research 257 (1989), S. 23-28 
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    ISSN: 1432-0878
    Keywords: Melatonin ; Immunohistochemistry ; Radioimmunoassay ; Pineal gland ; Mink, Mustela vison
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antiserum raised against N-amino-3-propyl melatonin bound to a protein carrier was used to visualize melatonin by immunohistochemistry and to measure melatonin concentration by radioimmunoassay in the pineal gland of intact mink females killed throughout the 24 h cycle and females killed after a bilateral ablation of the cervical superior ganglion. Melatonin immunoreactivity revealed by immunofluorescence or by the peroxidase-antiperoxidase complex was observed in the cytoplasm of presumed pinealocytes of all the females. Circadian changes in pineal melatonin content were not visualized by immunohistochemistry; furthermore, immunoreactivity was also present in the pineal gland of the ganglionectomized females. However, the melatonin content measured by radioimmunoassay was significantly higher in the pineal gland from intact females killed during the night compared with that of intact females killed during the day or of ganglionectomized females. The discrepancy between the results obtained using the two methods may arise because immunohistochemistry can detect very small amounts of melatonin.
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    Calbindin immunoreactivity in the neurons of the spinal cord and dorsal root ganglion of the domestic fowl (1989)
    Springer
    Cell & tissue research 257 (1989), S. 149-153 
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    ISSN: 1432-0878
    Keywords: Calbindin ; Brachial spinal cord ; Dorsal root ganglion ; Immunohistochemistry ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of the vitamin D-dependent calcium-binding protein, calbindin, was mapped in the brachial spinal cord and in the 15th dorsal root ganglion of the domestic fowl, using fluorescence immunohistochemistry. Cell somata of the dorsal root ganglion ranged in area from 200 μm2 to 2000 μm2. Sixteen percent of cell bodies displayed calbindin immunoreactivity. Reactivity occurred in both the small and large sensory neurons. These were randomly distributed within each ganglion. In the spinal cord, calbindin immunoreactivity was intense in Lissauer's tract, and in nerve fibres and nerve cell bodies within laminae 1 to 3 of the dorsal horn. Scattered varicose fibres were observed in laminae 4 to 7. Immunoreactivity was intense in laminae 10 where nerve fibres formed a meshwork around the central canal. Immunoreactive perikarya were occasionally observed in the outer region of lamina 10 and between laminae 8 and 9. The perikarya of the large motoneurons of lamina 9 were not reactive although they were enmeshed in calbindin-immunoreactive fibres.
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    URL: http://dx.doi.org/10.1007/BF00221645
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    Projections of neurons with neuromedin U-like immunoreactivity in the small intestine of the guinea-pig (1989)
    Springer
    Cell & tissue research 257 (1989), S. 415-422 
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    ISSN: 1432-0878
    Keywords: Neuromedin U ; Enteric nervous system ; Intestine, small ; Immunohistochemistry ; Neuropeptides ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neuromedin U immunoreactivity was located histochemically in the guinea-pig small intestine. Projections of immunoreactive neurons were determined by analysing patterns of degeneration following nerve lesions. The co-localization of neuromedin U immunoreactivity with immunoreactivity for substance P, neuropeptide Y, vasoactive intestinal peptide and calbindin was also investigated. Neuromedin U immunoreactivity was found in nerve cells in the myenteric and submucous plexuses and in nerve fibres in these ganglionated plexuses, around submucous arterioles and in the mucosa. Reactive fibres did not supply the muscle layers. Most reactive nerve cells in the myenteric ganglia had Dogiel type-II morphology and in many there was co-localization of calbindin, although some Dogiel type-II neuromedin U neurons were calbindin negative. Lesion studies suggest that these myenteric neurons project circumferentially to local myenteric ganglia. Projections from myenteric neurons also run anally in the myenteric plexus, while other projections extend to submucous ganglia, and still further projections run from the intestine to provide terminals in the coeliac ganglia. In the submucous ganglia neuromedin U was co-localized in three populations of nerve cells: (i) those with vasoactive intestinal peptide immunoreactivity, (ii) neurons containing neuropeptide Y, and (iii) neurons containing substance P. Each of these populations sends nerve fibres to the mucosa. Neuromedin U immunoreactivity is thus located in a variety of neurons serving different functions in the intestine and therefore probably does not have a single role in intestinal physiology.
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    Splenic outer periarterial lymphoid sheath (PALS): An immunoproliferative microenvironment constituted by antigen-laden marginal metallophils and ED2-positive macrophages in the rat (1989)
    Springer
    Cell & tissue research 257 (1989), S. 459-470 
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    ISSN: 1432-0878
    Keywords: Spleen ; Periarterial lymphoid sheath (PALS) ; Macrophage subpopulations ; Microenvironment ; B-lymphocytes ; Immune response ; Cell proliferation ; Immunohistochemistry ; Rat (DA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In an attempt to reveal the role of antigen-laden marginal metallophil (MM) and other macrophages in the intrasplenic immune response of a specific B-cell lineage to a thymus-independent type-2 antigen (Ficoll conjugated with fluorescein isothiocyanate), simultaneous immuno-histological observations of the involved cells were performed in the rat. By newly established methods of double or triple immunostainings, time-kinetics of the following parameters were studied and compared: (1) the antigen, (2) the specific antibody-forming cells (AFC) directed to the fluorescein-isothiocyanate determinant, (3) proliferating cells labeled with 5-bromo-2′-deoxyuridine (BrdU), and (4) macrophage subpopulations recognized by monoclonal antibodies (ED2 and ED3). The antigen localized stably not only in the marginal-zone macrophages but also in the MM except around the follicular area. The increase of BrdU-positive cells was observed from day 2 up to day 4 after antigen injection mostly in the periphery of the periarterial lymphoid sheath (outer PALS), which indicated antigen-induced proliferation. As a novel finding, the majority of AFC, both BrdU-positive and -negative, were either closely associated with the antigen-laden MM, or forming cell clusters with ED2-positive macrophages in the outer PALS. In contrast, there were very few AFC in juxtaposition to antigen-free MM in the follicular area or the antigen-laden marginal zone macrophages. The results led to the proposal of a hypothesis that the antigen-laden MM together with ED2-positive macrophages constitute an immunoproliferative microenvironment for the plasmacellular reaction by accumulating the antigen-specific B-cell lineage and promoting these cells to differentiate into the AFC and to proliferate in the outer PALS.
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    URL: http://dx.doi.org/10.1007/BF00221456
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    Immunocytochemical localization of hypocalcin in the endocrine cells of the corpuscles of Stannius in three teleost species (trout, flounder and goldfish) (1989)
    Springer
    Cell & tissue research 255 (1989), S. 651-656 
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    ISSN: 1432-0878
    Keywords: Corpuscles of Stannius ; Hypocalcin ; Immunohistochemistry ; Carassius auratus ; Hippoglossoides elassodon ; Salmo gairdneri
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In order to identify the cell-type responsible for the production of hypocalcin (the recently isolated hypocalcemic hormone of teleost fish), the corpuscles of Stannius (CS) of trout, flounder and goldfish, were immunocytochemically stained with antisera raised against trout hypocalcin. The secretory granules of the type-1 cells of the CS, considered to be the hypocalcin-producing cells, showed intense immunoreactivity in all species examined. However, in trout and flounder, the secretory granules produced by the type-2 cells, which have been suggested to represent a functionally different cell-type, also showed an intense immunoreactivity. In goldfish, no type-2 cells were observed. We tentatively conclude that type-1 and type-2 cells represent structurally different forms of the same functional cell-type.
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    Vasoactive intestinal polypeptide-like immunoreactivity in the bovine heart: high degree of coexistence with neuropeptide Y-like immunoreactivity (1989)
    Springer
    Cell & tissue research 256 (1989), S. 125-135 
    Details
    ISSN: 1432-0878
    Keywords: Heart-conducting system ; Vasoactive intestinal polypeptide (VIP) ; Neuropeptide Y ; Dopamine-betahydroxylase ; Immunohistochemistry ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary It is not known whether VIP coexists with other neuropeptides or neurotransmitters in the heart. In the present study, the distribution of nerve fibers showing VIP-like immunoreactivity (VIP-LI) in various parts of the bovine heart was compared with that of fibers possessing neuropeptide Y (NPY)- and dopamine-beta-hydroxylase (DBH)-LI. It was found that fibers showing VIP-LI most regularly occurred in the regions of sinuatrial and atrioventricular nodes and intracardiac ganglia, the nerve fibers being associated with nodal and ganglionic cells and arterial walls. Furthermore, it was observed that almost all fibers showing VIP-LI also exhibited NPY-LI. The fibers displaying VIP-LI occurred close to fibers endowed with DBH-LI, and on some occasions it appeared that varicosities showing immunolabeling for VIP also displayed DBH-LI. VIP- and NPY-LI, but not DBH-LI, was detected in some of the intracardiac ganglionic cells. The observations are discussed in relation to the previously known distribution and function of VIP in the heart and other organs. Further studies examining the effects of VIP and NPY in the heart should take into account possible interactions between these two peptides.
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    URL: http://dx.doi.org/10.1007/BF00224726
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    The relationship between gastrin cells and bombesin-like immunoreactive nerve fibres in the gastric antral mucosa of guinea-pig, rat, dog and man (1989)
    Springer
    Cell & tissue research 257 (1989), S. 171-178 
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    ISSN: 1432-0878
    Keywords: Gastrin ; Gastrin-releasing peptide ; Bombesin ; Stomach ; Autonomic innervation ; Immunohistochemistry ; Guinea pig ; Rat ; Dog ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The relationship between bombesin-like immunoreactive (bombesin-LI) nerve fibres and gastrin-LI G-cells was examined in gastric antral mucosa from guineapig, rat, dog and man using a double-labelling fluorescence immunohistochemical technique. The greatest density of bombesin-LI nerve fibres was found within the basal mucosa in all species and the density of innervation decreased towards the luminal surface. Most G-cells were in a band occupying approximately the middle third of the mucosa. The proportion of G-cells found within a distance of 2 μm from bombesin-LI nerve fibres was low in all species (6% in the guinea-pig, 22% in the rat, 14% in the dog, and 9% in the human). It is proposed that the neuropeptide released from bombesin-LI antral mucosal nerve fibres traverses distances of greater than several μm to reach the target G-cells. This may be achieved by passage through the mucosal microcirculation.
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    Immimohistochemical localization of corticotropin-releasing factor in selected brain areas of the European starling (Sturnus vulgaris) and the song sparrow (Melospiza melodia) (1989)
    Springer
    Cell & tissue research 257 (1989), S. 155-161 
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    ISSN: 1432-0878
    Keywords: Corticotropin-releasing factor (CRF) ; Immunohistochemistry ; Neurosecretory system ; Hypothalamus ; Nucleus striae terminalis ; European starling (Sturnus vulgaris) ; Song sparrow (Melospiza melodia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Corticotropin-releasing factor (CRF) was localized in the brains of two passerine species, the European starling (Sturnus vulgaris) and the song sparrow (Melospiza melodia), by means of immunohistochemistry. The hypothalamic distribution of this peptide in these species includes a complex of immunoreactive perikarya observed in the paraventricular nucleus (PVN), in both its medial and lateral divisions. Nerve fibers were also seen running from these areas to the anterior median eminence (AME) where a terminal field is apparent. A wide variety of extra-hypothalamic nuclei containing CRF-immunoreactive cells and fibers were identified. An apparent CRF terminal field can be visualized in the lateral septum. A dense fiber plexus is present in the nucleus accumbens (Ac) and more caudally in the nucleus of the stria terminalis (nST). In colchicinepretreated animals, it was revealed that these areas also contain CRF-stained perikarya. The pattern of CRF immunoreactivity in the Ac-nST complex is continuous, with no distinction apparent between the nuclei. The medial preoptic area (mPOA) and the adjacent diagonal band of Broca contain CRF-fibers, while cells are apparent in the mPOA. In the mesencephalon, cells were visualized in the midbrain central gray; a terminal field and scattered positively stained perikarya were found in areas more ventral to the central grey that are adjacent to the third cranial nerve. Scattered cells were also seen at the border of the nucleus intercollicularis-nucleus mesencephalicus lateralis, pars dorsalis complex. In contrast to mammalian studies, no immunoreactive nerve fibers or perikarya were observed in telencephalic areas homologous to the mammalian neocortex. These studies confirm the presence of a CRF path-way regulating pituitary function and suggest a broad role played by CRF as a neuromodulator or neurotransmitter in autonomic and possibly behavioral activities in these species.
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    Immunohistochemistry of vacuoles occurring in rat hepatocytes after retinol administration (1989)
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    Cell & tissue research 257 (1989), S. 263-268 
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    ISSN: 1432-0878
    Keywords: Retinol ; Vacuoles ; Immunohistochemistry ; Plasma proteins ; Hepatocytes ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The vacuoles occurring in rat hepatocytes after intraportal injection of retinol (33 or 67 μg) were examined immunohistochemically using respective antibodies against rat albumin, human retinol-binding protein, human ceruloplasmin, human α 1-antitrypsin, human transferrin, and human prealbumin as representative plasma proteins. The occurrence of the vacuoles reached a numerical maximum 30 min after injection of 67 μg retinol, followed by a temporal decrease. Hepatocytes from control rats, which had been intraportally injected with either blood plasma diluted to 2/3 concentration or with retinol palmitate solvent (castor oil) dissolved in blood plasma, showed immunoreactive fine granules without the occurrence of vacuoles in the cytoplasm. Identical vacuoles in serial sections appeared immunohistochemically either immunoreactive or non-immunoreactive for all the antibodies used, with rare exceptions. The occurrence of several rare exceptions suggested that 2 kinds of vacuoles might be formed in different cytoplasmic compartments. A zonal distribution of vacuoles was apparent in the hepatic laminae (or acini) within the liver lobules. The vacuoles were predominantly distributed in zone 2, and to a lesser extent in zone 3 and zone 1 in that order.
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    Localization and characterization of neuropeptide Y-like peptides in the brain and islet organ of the anglerfish (Lophius americanus) (1989)
    Springer
    Cell & tissue research 257 (1989), S. 303-311 
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    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Anglerfish peptide YG ; Brain ; Pancreas, endocrine, innervation ; Immunohistochemistry ; Radioimmunoassay ; High performance liquid chromatography ; Anglerfish, Lophius americanus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Results from a previous report demonstrate that more than one molecular form of neuropeptide Y-like peptide may be present in the islet organ of the anglerfish (Lophius americanus). Most of the neuropeptide Y-like immunoreactive material was anglerfish peptide YG, which is expressed in a subset of islet cells, whereas an additional neuropeptide Y-like peptide(s) was localized in islet nerves. To learn more about the neuropeptide Y-like peptides in islet nerves, we have employed immunohistochemical and biochemical methods to compare peptides found in anglerfish islets and brain. Using antisera that selectively react with either mammalian forms of neuropeptide Y or with anglerfish peptide YG, subsets of neurons were found in the brain that labelled with only one or the other of the antisera. In separate sections, other neurons that were labelled with either antiserum exhibited similar morphologies. Peptides from brains and islets were subjected to gel filtration and reverse-phase high performance liquid chromatography. Radioimmunoassays employing either the neuropeptide Y or peptide YG antisera were used to examine chromatographic eluates. Immunoreactive peptides having retention times of human neuropeptide Y and porcine neuropeptide Y were identified in extracts of both brain and islets. This indicates that peptides structurally similar to both of these peptides from the neuropeptide Y-pancreatic polypeptide family are expressed in neurons of anglerfish brain and nerve fibers of anglerfish islets. The predominant form of neuropeptide Y-like peptide in islets was anglerfish peptide YG. Neuropeptide Y-immunoreactive peptides from islet extracts that had chromatographic retention times identical to human neuropeptide Y and porcine neuropeptide Y were present in much smaller quantities. These results are consistent with the hypothesis that peptides having significant sequence homology with human neuropeptide Y and porcine neuropeptide Y are present in the nerve fibers that permeate the islet.
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    Reactivity of monoclonal islet cell antibodies on normal hyperplastic and neoplastic thyroid C-cells (1989)
    Springer
    Cell & tissue research 257 (1989), S. 437-443 
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    ISSN: 1432-0878
    Keywords: Monoclonal islet antibodies ; Thyroid C-cells ; Differentiation antigens ; Immunohistochemistry ; Cow, Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Thyroid C-cell reactivity to 15 monoclonal antibodies raised against a series of pancreatic islet cells (H[human]ISL, B[bovine]ISL and R[rat]ISL) was evaluated using an indirect immunoperoxidase technique on frozen thyroid sections. Of the monoclonal anti-islet cell antibodies, five reacted specifically with bovine C-cells or human hyperplastic and neoplastic C-cells but not with follicular cells. Two monoclonal antibodies of the bovine series showed strong immunoreactivity with C-cells and only a weakly positive immunostaining of follicular cells. Five monoclonal antibodies reacted with both thyroid C-cells and follicular cells, whereas 3 monoclonal anti-islet cell antibodies did not stain any cell type of the thyroid. In human medullary carcinomas, calcitonin- and somatostatin-producing neoplastic cells were immunoreactive with the same monoclonal antibodies as were normal human C-cells. The protein bands identified by the monoclonal antibodies in human medullary carcinomas had the same molecular weight as those from pancreatic islet extracts. Our study demonstrates the presence of similar differentiation antigens on thyroid C-cells and pancreatic islet cells; this further illustrates common modes of differentiation and specialisation of these embryologically different members of the dispersed neuroendocrine system. The crossreactivity of seven of the monoclonal antibodies investigated with follicular epithelium of the thyroid suggests the existence of common antigenic determinants in different endocrine organs and may partly explain the multiple organ autoimmune response found in patients with polyendocrine diseases.
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    Immunogold localisation of endogenous immunoglobulin-G in ultrathin frozen sections of the human placenta (1989)
    Springer
    Cell & tissue research 257 (1989), S. 603-607 
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    ISSN: 1432-0878
    Keywords: Placenta ; Immunoglobulin G ; Endocytosis ; Immunohistochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Endogenous immunoglobulin-G was localised in ultrathin frozen sections of human term placenta by use of an indirect immuno electron-histochemical methodology. Immunoreactivity of endogenous IgG to rabbit anti-human immunoglobulin-G antibody was visualised by use of protein-A — colloidal gold complex. Gold marked the syncytiotrophoblast in both coated and uncoated regions of the apical plasmalemma, in vesicles and multivesicular bodies, and in vesicles near the basal plasmalemma. Immunoreactivity was also seen in the interstitial space between the trophoblast and the fetal endothelial layer as well as in various types of vesicles within the endothelial cells. No immunoreactivity was seen in the intercellular clefts of the endothelium. The pattern of localisation observed is consistent with receptor-mediated uptake of immunoglobulin-G into the syncytiotrophoblast of the human placenta followed by release into the interstitial space and then vesciular transport through the endothelium.
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    The occurrence and distribution of peptide- or 5-HT-containing nerves in the swimbladder of four different species of teleosts (Gadus morhua, Ctenolabrus rupestris, Anguilla anguilla, Salmo gairdneri) (1989)
    Springer
    Cell & tissue research 257 (1989), S. 641-647 
    Details
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Immunohistochemistry ; Swimbladder ; Gas gland ; Gadus morhua ; Ctenolabrus rupestris ; Anguilla anguilla ; Salmo gairdneri (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The innervation of the swimbladder in four different teleost species has been studied by the use of immunohistochemical methods. The teleosts examined belong to two different groups regarding their swimbladder morphology: physoclists (the cod, Gadus morhua and the goldsinny wrasse, Ctenolabrus rupestris) and physostomes (the eel, Anguilla anguilla and the rainbow trout, Salmo gairdneri). Vasoactive intestinal polypeptide-like immunoreactivity was demonstrated in nerves of the swimbladder walls of all four species, and in the gas glands of the cod and the goldsinny wrasse. Substance P-like immunoreactivity was shown in swimbladders of the cod, eel and rainbow trout but not the goldsinny wrasse. Immunoreactivity to met-enkephalin antiserum was revealed in the swimbladder walls of the eel and the goldsinny wrasse, while neurotensin-like immunoreactivity was present in the goldsinny wrasse and rainbow trout swimbladders. Neurotensin-like immunoreactivity was also seen in the gas gland of the goldsinny wrasse. 5-Hydroxytryptamine immunoreactivity was found in endocrine cells in the pneumatic duct of the eel and in the swimbladder walls of the goldsinny wrasse and the rainbow trout. In conclusion, all teleosts examined showed a very close resemblance in the peptidergic/tryptaminergic innervation of the swimbladder to that of the gut, inasmuch as the immunoreactivity present in the swimbladders always occurred in the gut of the same species.
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    Dependence of inessential late gene expression on early meiotic events in Saccharomyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 215 (1989), S. 490-500 
    Details
    ISSN: 1617-4623
    Keywords: Meiosis ; Saccharomyces cerevisiae ; Sporulation ; Inessential genes ; Meiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary SPR3 is one of at least nine genes which are expressed in sporulating Saccharomyces cerevisiae cells at the time of meiosis I. We show below that strains homozygous for null alleles of SPR3 are capable of normal meiosis and the production of viable ascospores. We have also monitored SPR3 expression in a series of strains that are defective in meiotic development, using an SPR3: lacZ fusion carried on a single copy plasmid. β-Galactosidase activity occurred at wild-type levels in diploid strains homozygous for mutations in spo13, rad50, rad57 and cdc9, but was greatly reduced in strains carrying cdc8 or spo7 defects. We conclude that SPR3 expression is a valid monitor of early meiotic development, even though the gene is inessential for the sporulation process.
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    Structure of the HOM2 gene of Saccharomyces cerevisiae and regulation of its expression (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 149-154 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; HOM2 gene ; Aspartic semi-aldehyde ; General control
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Saccharomyces cerevisiae the HOM2 gene encodes aspartic semi-aldehyde dehydrogenase (ASA DH). The synthesis of this enzyme had been shown to be derepressed by growth in the presence of high concentrations of methionine. In the present work we have cloned and sequenced the HOM2 gene and found that the promoter region of this gene bears one copy of the consensus sequence for general control of amino acid synthesis. This prompted us to study the regulation of the expression of the HOM2 gene. We have found that ASA DH is the first reported enzyme of the related threonine and methionine pathway to be regulated by the general control of amino acid synthesis.
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    Chromatin structure of the 5′ flanking region of the yeastLEU2 gene (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 464-470 
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    ISSN: 1617-4623
    Keywords: Nucleosome positioning ; LEU2 gene ; Saccharomyces cerevisiae ; tRNA3 Leu gene ; Chromatin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The chromatin structure of theLEU2 gene and its flanks has been studied by means of nuclease digestion, both with micrococcal nuclease and DNase I. The gene is organized in an array of positioned nucleosomes. Within the promoter region, the nucleosome positioning places the regulatory sequences, putative TATA box and upstream activator sequence outside the nucleosomal cores. The tRNA3 Leu gene possesses a characteristic structure and is protected against nucleases. Most of the 5′ flank is sensitive to DNase I digestion, although no clear hypersensitive sites were found. The chromatin structure is independent of either the transcriptional state of the gene or the chromosomal or episomal location. Finally, in the plasmid pJDB207, which lacks most of the promoter, we have found that the chromatin structure of the coding region is similar to that of the wild-type allele.
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    The naturally occurring silent invertase structural gene suc2° contains an amber stop codon that is occasionally read through (1989)
    Springer
    Molecular genetics and genomics 216 (1989), S. 511-516 
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    ISSN: 1617-4623
    Keywords: Nonsense mutation ; Read-through ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The yeast invertase structural gene SUC2 has two naturally occurring alleles, the active one and a silent allele called suc2°. Strains carrying suc2° are unable to ferment sucrose and do not show detectable invertase activity. We have isolated suc2° and found an amber codon at position 232 of 532 amino acids. However, transformants carrying suc2° on a multicopy plasmid were able to ferment sucrose and showed detectable invertase activity. Full-length invertase was found in gels stained for active invertase and in immunoblots. Therefore we concluded that the amber codon is occasionally read as an amino acid. The calculated frequency of read-through is about 4% of all translation events.
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    URL: http://dx.doi.org/10.1007/BF00334398
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    Expression and DNA sequence of RED1, a gene required for meiosis I chromosome segregation in yeast (1989)
    Springer
    Molecular genetics and genomics 218 (1989), S. 293-301 
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    ISSN: 1617-4623
    Keywords: Sporulation ; DNA sequence ; Saccharomyces cerevisiae ; Meiosis ; Chromosome segregation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Genetic studies have previously demonstrated that the RED1 gene of Saccharomyces cerevisiae is required for chromosome segregation at the first meiotic division. Northern blot hybridization analysis indicates that the RED1 gene produces two transcripts of 2.75 and 3.2 kilobases. The major 2.75 kb transcript is not present in mitotic cells and is meiotically induced to accumulate maximally just prior to the meiosis I division. The DNA sequence of the RED1 gene was determined and used to predict the amino acid sequence of the encoded gene product. The RED1 protein is 827 amino acids in length and has a molecular weight of 95.5 kilodaltons. There is no significant homology between the RED1 amino acid sequence and other known protein sequences, including those encoded by genes essential for meiosis.
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    URL: http://dx.doi.org/10.1007/BF00331281
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    Mutant invertase proteins accumulate in the yeast endoplasmic reticulum (1989)
    Springer
    Molecular genetics and genomics 215 (1989), S. 401-406 
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    ISSN: 1617-4623
    Keywords: Protein secretion ; Saccharomyces cerevisiae ; Invertase ; Endoplasmic reticulum ; Secretion mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Intercompartmental transport of secreted proteins in yeast was analysed using invertase mutants. Deletions and insertions at the BamHI (position +787) or the Asp718 (position +1159) sites of the SUC2 gene led to mutant proteins with different behaviour regarding secretion, localization and enzyme activity. The deletion mutants showed accumulation of core glycosylated material in the endoplasmic reticulum (ER) a decrease of secreted protein by 5%–30% and loss of enzyme activity. The secreted material was localized in the culture medium and not — as is normal for invertase-in the cell wall. No delay in transport from the Golgi to the cell surface was observed, indicating that the rate-limiting step for secretion is at the ER-Golgi stage. Two insertion mutants, pIPA and pIPB, retained enzyme activity. Mutant pIPB showed 10% secretion, while 60%–70% secretion was observed for pIPA. While the non-secreted material accumulated in the ER, the secreted material was present in the cell wall. The results suggest that the presence of structures incompatible with secretion leads to ER accumulation of mutated invertase.
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    URL: http://dx.doi.org/10.1007/BF00427036
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    The bI4 RNA mitochondrial maturase of Saccharomyces cerevisiae can stimulate intra-chromosomal recombination in Escherichia coli (1989)
    Springer
    Molecular genetics and genomics 216 (1989), S. 70-74 
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    ISSN: 1617-4623
    Keywords: RNA splicing ; Maturase ; Recombinase ; Mitochondria ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary When the bI4 RNA maturase, encoded by the fourth intron of the mitochondrial cytochrome b gene of Saccharomyces cerevisiae, was expressed in Escherichia coli, formation of intra-chromosomal Lac+ recombinants was stimulated threefold. This “hyper-rec” phenotype was recA as well as recBCD dependent. The most active form of the bI4 maturase stimulated homologous recombination whereas splicing deficient mutants of bI4 maturase were either deficient in or unable to stimulate homologous recombination.
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    In vitro and in vivo studies on the mitochondrial import of CBS1, a translational activator of cytochrome b in yeast (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 162-167 
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    ISSN: 1617-4623
    Keywords: Nucleotide sequence ; PET gene ; Mitochondrial import ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Translation of mitochondrial cytochrome b mRNA in yeast is activated by the product of the nuclear gene CBS1. CBS1 encodes a 27 kDa precursor protein, which is cleaved to a 24 kDa mature protein during the import into isolated mitochondria. The sequences required for mitochondrial import reside in the amino-terminal end of the CBS1 precursor. Deletion of the 76 amino-terminal amino acids renders the protein incompetent for mitochondrial import in vitro and non-functional in vivo. When present on a high copy number plasmid and under the control of a strong yeast promoter, biological function can be restored by this truncated derivative. This observation indicates that the CBS1 protein devoid of mitochondrial targeting sequences can enter mitochondria in vivo, possibly due to a bypass of the mitochondrial import system.
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    Mitochondrial introns aI1 and/or aI2 are needed for the in vivo deletion of intervening sequences (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 168-171 
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    ISSN: 1617-4623
    Keywords: Mitochondrial introns ; Reverse transcriptase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Some pet- (or mit-) mutations impeding the splicing of one or several intron(s) of the yeast mitochondrial pre-mRNA(s) are suppressed in vivo by the DNA deletion of these introns. We have genetically demonstrated that introns aI1 and/or aI2 of the cytochrome c oxidase subunit 1 gene are necessary for this deletion process. The facts that adjacent introns are simultaneously deleted and that, in the pet- (or mit-) mutants which easily revert by intron deletion, the splicing of the introns they affect is only partially blocked, suggest that the intron encoded proteins aI1 and/or aI2 could intervene by means of their putative reverse transcriptase activity.
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    Reassembly of microtubules in protoplasts ofSaccharomyces cerevisiae after nocodazole-induced depolymerization (1989)
    Springer
    Protoplasma 151 (1989), S. 98-105 
    Details
    ISSN: 1615-6102
    Keywords: Microtubule neoformation ; Nocodazole ; Protoplasts ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary By following microtubule neoformation after their complete destruction by nocodazole, we analyzed the pattern of microtubule nucleation in protoplasts ofSaccharomyces cerevisiae. Using immunofluorescence, the drug was shown to induce rapid and complete disassembly of both cytoplasmic and spindle microtubules and to selectively block protoplast nuclear division at a defined stage of the cell cycle. Treated protoplasts placed in a drug-free environment recovered a more abundant microtubular system. The majority of microtubules re-formed at SPBs whereas a minority of free-ended microtubules nucleated in the cytoplasm of the protoplasts without any detectable association with recognizable nucleation sites. Random nucleation of free microtubules might be induced by high amounts of unpolymerized tubulin likely to be present in the protoplasts at the moment of drug release.
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    Analysis of expression of hybrid yeast genes containing ARS elements (1989)
    Springer
    Molecular genetics and genomics 218 (1989), S. 531-535 
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    ISSN: 1617-4623
    Keywords: Origin of replication ; Promoter ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In an attempt to devise a new assay for ARS-binding proteins we have inserted the HO ARS between the upstream activation site and the TATA region of the yeast CYC1 promoter. A marked reduction in promoter activity is observed. Inactivation of the HO ARS element by point mutation does not restore promoter activity to its original level, although a modest activation is seen. We have also inserted the HO ARS into the intron of the yeast actin gene; although there is no apparent deleterious effect on transcription, the activity of the ARS is abolished in this new environment.
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    Identification of a gene conferring resistance to zinc and cadmium ions in the yeast Saccharomyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 219 (1989), S. 161-167 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Heavy metal resistance ; DNA sequence ; Membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A DNA fragment conferring resistance to zinc and cadmium ions in the yeast Saccharomyces cerevisiae was isolated from a library of yeast genomic DNA. Its nucleotide sequence revealed the presence of a single open reading frame (ORF; 1326 bp) having the potential to encode a protein of 442 amino acid residues (molecular mass of 48.3 kDa). A frameshift mutation introduced within the ORF abolished resistance to heavy metal ions, indicating the ORF is required for resistance. Therefore, we termed it the ZRC1 (zinc resistance conferring) gene. The deduced amino acid sequence of the gene product predicts a rather hydrophobic protein with six possible membrane-spanning regions. While multiple copies of the ZRC1 gene enable yeast cells to grow in the presence of 40 mM Zn2+, a level at which wild-type cells cannot survive, the disruption of the chromosomal ZRC1 locus, though not a lethal event, makes cells more sensitive to zinc ions than are wild-type cells.
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    Isolation and characterization of Saccharomyces cerevisiae mutants supersensitive to G1 arrest by the mating hormone a-factor (1989)
    Springer
    Molecular genetics and genomics 219 (1989), S. 439-444 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; a-factor ; Conjugation ; G1 arrest ; ssl mutations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Nine independent mutants which are supersensitive (ssl −) to G1 arrest by the mating hormone a-factor were isolated by screening mutagenized Saccharomyces cerevisiae MATα cells on solid medium for increased growth inhibition with a-factor. These mutants carried lesions in two complementation groups, ssl1 and ssl2. Mutations at the ssl1 locus were mating type specific: MATα ssl1 − cells were supersensitive to α-factor but MATα ssl1 − were not supersensitive to α-factor. In contrast, mutations at the ssl2. locus conferred supersensitivity to the mating hormone of the opposite mating type on both MATα, and MATa cells. The α-cell specific capacity to inactivate externally added a-factor was shown to be lacking in MATα ssl1 − mutants whereas MATα ssl2. cells were able to inactivate a-factor. Complementation analysis showed that ssl2 and sst2, a mutation originally isolated as conferring supersensitivity to α-factor to MATa cells, are lesions in the same gene. The ssl1 gene was mapped 30.5 centi-Morgans distal to ilv5 on chromosome XII.
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    A full length cDNA clone for the alkaline protease from Aspergillus oryzae: Structural analysis and expression in Saccharomyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 219 (1989), S. 33-38 
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    ISSN: 1617-4623
    Keywords: Aspergillus oryzae ; Alkaline protease ; Prepro sequence ; Heterologous expression ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned and determined the nucleotide sequence of a cDNA fragment for the entire coding region of the alkaline protease (Alp) from a filamentous ascomycete Aspergillus oryzae. According to the deduced amino acid sequence, Alp has a putative prepro region of 121 amino acids preceding the mature region, which consists of 282 amino acids. A consensus sequence of a signal peptide consiting of 21 amino acids is found at the N-terminus of the prepro region. The primary structure of the mature region shares extensive homology (29%–44%) with those of subtilisin families, and the three residues (Asp 32, His 64 and Ser 221 in subtilisin BPN′) composing the active site are preserved. The entire cDNA, coding for prepro Alp, when introduced into the yeast Saccharomyces cerevisiae, directed the secretion of enzymatically active Alp into the culture medium, with its N-terminus and specific activity identical to native Aspergillus Alp.
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    The PS03 gene is involved in error-prone intragenic recombinational DNA repair in Saccharomyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 219 (1989), S. 75-80 
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    ISSN: 1617-4623
    Keywords: Gene conversion ; Crossing-over ; Mismatch repair ; Saccharomyces cerevisiae ; Psoralens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The induction of gene conversion and mitotic crossing-over by photoaddition of psoralens, 254 nm ultraviolet radiation, and nitrogen mustards was determined in diploid cells homozygous for the pso3-1 mutation and in the corresponding wild type of Saccharomyces cerevisiae. For these different agents, the frequency of non-reciprocal events (conversion) is reduced in the pso3-1 mutant compared to the wild type. In contrast, the frequency of reciprocal events (crossing-over) is increased at a range of doses. These observations, together with the block in induced mutagenesis for both reverse and forward mutations previously reported for the pso3-1 mutant, suggest that the PS03 gene product plays a role in mismatch repair of short patch regions. The block in gene conversion in the pso3 homozygous diploid leads, in the case of nitrogen mustards, to specific repair intermediates which are lethal to the cells.
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    Yeast mutants with enhanced ability to secrete human lysozyme: Isolation and identification of a protease-deficient mutant (1989)
    Springer
    Molecular genetics and genomics 219 (1989), S. 58-64 
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    ISSN: 1617-4623
    Keywords: Enhanced secretion ; Human lysozyme production ; Protease mutant ; Protein processing ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Yeast mutant strains which secrete large amounts of human lysozyme were screened using an agar medium containing bacterial cells. Nine mutants secreted over 10 times more lysozyme than the wild-type parent strain. The mRNA levels for lysozyme in the mutants were not higher than that of the wild-type strain. Three of the mutant strains were deficient in carboxypeptidase Y activity. It was found that the protease deficiency was caused by a deficiency in conversion of proenzyme to mature enzyme in ssl1 mutant cells. The ssl1 gene was found to be closely linked to the centromere and determine both the efficiency of secretion of lysozyme and the processing of carboxypeptidase Y.
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    Plasmid multimerization is dependent on RAD52 activity in Saccharomyces cerevisiae (1989)
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    Molecular genetics and genomics 219 (1989), S. 495-498 
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    ISSN: 1617-4623
    Keywords: ARS1 ; Plasmid multimerization ; RAD52 ; Saccharomyces cerevisiae ; Eukaryotic recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A mutant plasmid, pX, derived from the 1453 base pair small plasmid, YARp1 (or TRP1 RI circle), consists of 849 base pairs of DNA bearing the TRP1 gene and the ARS1 sequence of Saccharomyces cerevisiae and, unlike YARp1 and other commonly used yeast plasmids, highly multimerizes in a S. cerevisiae host. The multimerization of pX was dependent on RAD52, which is known to be necessary for homologous recombination in S. cerevisiae. Based upon this observation, a regulated system of multimerization of pX with GAL1 promoter-driven RAD52 has been developed. We conclude that the regulated multimerization of pX could provide a useful model system to study genetic recombination in the eukaryotic cell, in particular to investigate recombination intermediates and the effects of various trans-acting mutations on the multimerization and recombination of plasmids.
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    Structure and expression of the URA5 gene of Saccharomyces cerevisiae (1989)
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    Molecular genetics and genomics 215 (1989), S. 455-462 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; Orotate phosphoribosyl transferase ; Nucleotide sequence ; Transcription ; Regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The URA5 gene of Saccharomyces cerevisiae encodes orotate phosphoribosyl transferase (EC 2.4.2.10; OPRTase) which catalyses the transformation of orotate to OMP in the pyrimidine pathway. We present in this paper the cloning and the sequencing of this gene, the last in the yeast pyrimidine pathway to be cloned. We have deduced the protein sequence of the OPRTase of S. cerevisiae from the DNA sequence and compared it to that of Escherichia coli, Podospora anserina and Dictyostelium discoideum. Some important similarities in the structure of these four proteins have been found. Finally, we have quantified the transcription of the URA5 gene in different physiological conditions and confirmed that it was not under the control of UTP or any intermediary product of the pathway.
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    Accumulation of the cytochrome c oxidase subunits I and II in yeast requires a mitochondrial membrane-associated protein, encoded by the nuclear SCO1 gene (1989)
    Springer
    Molecular genetics and genomics 216 (1989), S. 37-43 
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    ISSN: 1617-4623
    Keywords: DNA sequence ; PET gene ; Saccharomyces cerevisiae ; Mitochondrial import ; cytochrome c oxidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The yeast nuclear SCO1 gene is required for accumulation of the mitochondrially synthesized cytochrome c oxidase subunits I and II (COXI and COXII). We cloned and characterized the SCO1 gene. It codes for a 0.9 kb transcript. DNA sequence analysis predicts a 33 kDa protein. As shown by in vitro transcription and translation experiments in combination with import studies on isolated mitochodria, this protein is matured into a 30 kDa polypeptide which is tightly associated with a mitochondrial membrane. The possible function of the SCO1 gene product in the assembly of cytochrome c oxidase is discussed.
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    Three regulatory systems control expression of glutamine synthetase inSaccharomyces cerevisiae at the level of transcription (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 370-377 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; GLN1 ; Glutamine synthetase ; Regulatory systems ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary TheGLN1 gene ofSaccharomyces cerivisiae was cloned by complementation of agln1 auxotroph. AGLN1-lacZ fusion was constructed to assayGLN1 promoter activity. β-Galactosidase and glutamine synthetase expression in chromosomally integratedGLN1-lacZ fusion strains were co-regulated in response to a shift from glutamine to glutamate as the nitrogen source, purine limitation, and 3-aminotriazole-induced histidine starvation. Regulation ofGLN1 expression by each of the three pathways occurred at the transcriptional level. Increased accumulation ofGLN1 mRNA was observed within 5 min after a shift from glutamine to glutamate as the nitrogen source. After 5 min following glutamine addition to the cells growing with glutamate as nitrogen source. This indicates that theGLN1 message is unstable and has a half-life of approximately 3 min. Deletion analysis indicated that the sequences required forGLN1 expression are located within approximately 350 bp upstream from the transcriptional initiation site.
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    Molecular cloning of SNM1, a yeast gene responsible for a specific step in the repair of cross-linked DNA (1989)
    Springer
    Molecular genetics and genomics 218 (1989), S. 64-71 
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    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; DNA repair ; Cross-link ; Transposon mapping ; Nitrogen mustard
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated yeast gene SNM1 via complementation of sensitivity towards bi- and tri-functional alkylating agents in haploid and diploid yeast DNA repair-deficient snm1-1 mutants. Four independent clones of plasmid DNA containing the SNM1 locus were isolated after transformation with a YEp24-based yeast gene bank. Subcloned SNM1-containing DNA showed (i) complementation of the repair-deficiency phenotype caused by either one of the two different mutant alleles snm1-1 and snm1-2 ts; (ii) complementation in haploid and diploid yeast snm1-1 mutants by either single or multiple copies of the SNM1 locus; and (iii) that the SNM1 gene is at most 2.4 kb in size. Expression of SNM1 on the smallest subclone, however, was under the control of the GAL1 promotor. Gene size and direction of transcription was further verified by mutagenesis of SNM1 by Tn10-LUK transposon insertion. Five plasmids containing Tn10-LUK insertions at different sites of the SNM1-containing DNA were able to disrupt function of genomic SNM1 after gene transplacement. Correct integration of the disrupted SNM1::Tn10-LUK at the genomic site of SNM1 was verified via tetrad analysis of the sporulated diploid obtained after mating of the SNM1::Tn10-LUK transformant to a haploid strain containing the URA3 SNM1 wild-type alleles. The size of the poly(A)+ RNA transcript of the SNM1 gene is 1.1 kb as determined by Northern analysis.
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    URL: http://dx.doi.org/10.1007/BF00330566
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    ThePSO4 gene is responsible for an error-prone recombinational DNA repair pathway inSaccharomyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 419-426 
    Details
    ISSN: 1617-4623
    Keywords: pso4-1 mutation ; Saccharomyces cerevisiae ; Error-prone recombinational repair ; Mitotic recombination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The induction of mitotic gene conversion and crossing-over inSaccharomyces cerevisiae diploid cells homozygous for thepso4-1 mutation was examined in comparison to the corresponding wild-type strain. Thepso4-1 mutant strain was found to be completely blocked in mitotic recombination induced by photoaddition of mono- and bifunctional psoralen derivatives as well as by mono- (HN1) and bifunctional (HN2) nitrogen mustards or 254 nm UV radiation in both stationary and exponential phases of growth. Concerning the lethal effect, diploids homozygous for thepso4-1 mutation are more sensitive to all agents tested in any growth phase. However, this effect is more pronounced in the G2 phase of the cell cycle. These results imply that the ploidy effect and the resistance of budding cells are under the control of thePSO4 gene. On the other hand, thepso4-1 mutant is mutationally defective for all agents used. Therefore, thepso4-1 mutant has a generalized block in both recombination and mutation ability. This indicates that thePSO4 gene is involved in an error-prone repair pathway which relies on a recombinational mechanism, strongly suggesting an analogy between thepso4-1 mutation and theRecA orLexA mutation ofEscherichia coli.
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    URL: http://dx.doi.org/10.1007/BF02464912
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    Putative target sites for mobile G+C rich clusters in yeast mitochondrial DNA: Single elements and tandem arrays (1989)
    Springer
    Molecular genetics and genomics 218 (1989), S. 272-283 
    Details
    ISSN: 1617-4623
    Keywords: GC clusters ; Mobile elements ; Target sites ; mtDNA ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary GC clusters constitute the major repetitive elements in the mitochondrial (mt) genome of the yeast Saccharomyces cerevisiae. Many of these clusters are optional and thus contribute much to the polymorphism of yeast mtDNAs. We have made a systematic search for polymorphic sites by comparing mtDNA sequences of various yeast strains. Most of the 26 di- or polymorphic sites found differ by the presence or absence of a GC cluster of the majority class, here referred to as the M class, which terminate with an AGGAG motif. Comparison of sequences with and without the GC clusters reveal that elements of the subclasses M1 and M2 are inserted 3′ to a TAG, flanked by A+T rich sequences. M3 elements, in contrast, only occur in tandem arrays of two to four GC clusters; they are consistently inserted 3′ to the AGGAG terminal sequence of a preexisting cluster. The TAG or the terminal AGGAG, therefore, are regarded as being part of the target sites for M1 and M2 or M3 elements, respectively. The dinucleotide AG is in common to both target sites; it also occurs at the 3′ terminus (AGGAG). This suggests its duplication during GC cluster insertion. This notion is supported by the observation that GC clusters of the minor classes G and V similarily repeat at their 3′ terminus a GT or an AA dinucleotide, respectively, from their putative target sites.
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    URL: http://dx.doi.org/10.1007/BF00331278
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    Chromosomal localization and expression of CBS1, a translational activator of cytochrome b in yeast (1989)
    Springer
    Molecular genetics and genomics 218 (1989), S. 57-63 
    Details
    ISSN: 1617-4623
    Keywords: Saccharomyces cerevisiae ; PET gene ; Transcriptional regulation ; Anaerobiosis ; 5′ mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Translation of mitochondrial cytochrome b RNA in yeast requires the product of the nuclear gene CBS1, a 27.5 kDa soluble mitochondrial protein. In this paper we show that the CBS1 gene is located on chromosome IV immediately adjacent to COX9, the gene coding for cytochrome c oxidase subunit VIIa. CBS1 is transcribed as a very low abundant 900 b RNA. Transcription starts at a single position 101 bp upstream of the CBS1 initiation codon. At positions-39 to-27 of its leader sequence it contains a small open reading frame of 4 codons. By monitoring the β-galactosidase activity of a CBS1/lacZ fusion construct we show that expression of CBS1 is subjected to regulation by oxygen and by glucose: the β-galactosidase activity is elevated threefold in glycerol or galactose grown cells compared to that in glucose grown cells. A further threefold reduction of the activity is observed in anaerobically grown cells. In accordance with this result is the observation that the steady-state level of CBS1 mRNA of anaerobically grown cells is ninefold lower than that of aerobically cultured cells.
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    URL: http://dx.doi.org/10.1007/BF00330565
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    Structure and function of the yeast vacuolar membrane proton ATPase (1989)
    Springer
    Journal of bioenergetics and biomembranes 21 (1989), S. 589-603 
    Details
    ISSN: 1573-6881
    Keywords: Vacuolar membrane H+ATPase ; vacuoles ; Saccharomyces cerevisiae ; catalytic cooperativity of ATP hydrolysis ; VMA genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Our current work on a vacuolar membrane proton ATPase in the yeastSaccharomyces cerevisiae has revealed that it is a third type of H+-translocating ATPase in the organism. A three-subunit ATPase, which has been purified to near homogeneity from vacuolar membrane vesicles, shares with the native, membrane-bound enzyme common enzymological properties of substrate specificities and inhibitor sensitivities and are clearly distinct from two established types of proton ATPase, the mitochondrial F0F1-type ATP synthase and the plasma membrane E1E2-type H+-ATPase. The vacuolar membrane H+-ATPase is composed of three major subunits, subunita (M r =67 kDa),b (57kDa), andc (20 kDa). Subunita is the catalytic site and subunitc functions as a channel for proton translocation in the enzyme complex. The function of subunitb has not yet been identified. The functional molecular masses of the H+-ATPase under two kinetic conditions have been determined to be 0.9–1.1×105 daltons for single-cycle hydrolysis of ATP and 4.1–5.3×105 daltons for multicycle hydrolysis of ATP, respectively.N,N′-Dicyclohexylcarbodiimide does not inhibit the former reaction but strongly inhibits the latter reaction. The kinetics of single-cycle hydrolysis of ATP indicates the formation of an enzyme-ATP complex and subsequent hydrolysis of the bound ATP to ADP and Pi at a 7-chloro-4-nitrobenzo-2-oxa-1,3-diazolesensitive catalytic site. Cloning of structural genes for the three subunits of the H+-ATPase (VMA1, VMA2, andVMA3) and their nucleotide sequence determination have been accomplished, which provide greater advantages for molecular biological studies on the structure-function relationship and biogenesis of the enzyme complex. Bioenergetic aspects of the vacuole as a main, acidic compartment ensuring ionic homeostasis in the cytosol have been described.
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    URL: http://dx.doi.org/10.1007/BF00808115
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    Molecular properties of the fungal plasma-membrane [H+]-ATPase (1989)
    Springer
    Journal of bioenergetics and biomembranes 21 (1989), S. 621-632 
    Details
    ISSN: 1573-6881
    Keywords: ATPase ; [H+]-ATPase ; proton transport ; Neurospora crassa ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract The fungal plasma membrane contains a proton-translocating ATPase that is closely related, both structurally and functionally, to the [Na+, K+]-, [H+, K+]-, and [Ca2+]-ATPases of animal cells, the plasma-membrane [H+]-ATPase of higher plants, and several bacterial cation-transporting ATPases. This review summarizes currently available information on the molecular genetics, protein structure, and reaction cycle of the fungal enzyme. Recent efforts to dissect structure-function relationships are also discussed.
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    URL: http://dx.doi.org/10.1007/BF00808117
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    High frequency FLP-independent homologous DNA recombination of 2 μ plasmid in the yeast Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 14 (1988), S. 191-199 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Site specific recombination ; 2 μ DNA plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The purpose of this work is to identify and quantitate in vivo 2 μ plasmid FLP-independent recombination in yeast, using a nonselective assay system for rapid detection of phenotypic expression of the recombination events. A tester plasmid was constructed such that in vivo recombination between 2 μ direct repeat sequences produces the resolution of the plasmid into two circular DNA molecules. This recombinational event is detected as a phenotypic shift from red to white colonies, due to the mitotic loss of the plasmid portion containing the yeast ADE8 gene in a recipient ade1 ade2 ade8 genetic background. In the absence of the 2 μ FLP recombinase and/or its target DNA sequence, recombination is not abolished but rather continues at a high frequency of about 17%. This suggests that the FLP-independent events are mediated by the chromosomally-encoded general homologous recombination system. We therefore conclude that the totality of 2 μ DNA recombination events occurring in FLP+ cells is the contribution of both FLP-mediated and FLP-independent events.
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    URL: http://dx.doi.org/10.1007/BF00376739
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    Molecular cloning of the ARG7 gene of Schizosaccharomyces pombe encoding argininosuccinate lyase (1988)
    Springer
    Current genetics 14 (1988), S. 381-385 
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    ISSN: 1432-0983
    Keywords: Schizosaccharomyces pombe ; Saccharomyces cerevisiae ; Argininosuccinate lyase ; Gene cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A gene bank of Sau3A partially restricted Schizosaccharomyces pombe DNA in YEp13 was used to transform an arg4 mutant of Saccharomyces cerevisiae. One colony was recovered which contained the YEp13 plasmid bearing a large insert complementing the argininosuccinate lyase (ASL) mutation. As shown by restriction mapping and subcloning experiments, the DNA sequence required for complementation is localized on a 2 kb BamHI-BamHI fragment. The plasmid complemented several S. cerevisiae arg4 mutants of independent origin and a S. pombe arg7 mutant lacking ASL. Low but significant ASL activities were detected in crude extracts of these transformants. No complementation of the E. coli argH mutant was observed. Southern blot hybridizations showed that the insert originates from the S. pombe genome. No cross-hybridization was found between this sequence and S. cerevisiae DNA. It can be concluded that the cloned DNA fragment bears the S. pombe ARG7 gene coding for ASL.
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    Inheritance of chromosome length polymorphisms in Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 14 (1988), S. 413-418 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Chromosome length polymorphisms ; FIGE ; OFAGE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Although Saccharomyces cerevisiae strains generally have similar chromosomal band patterns as revealed by pulsed field gel electrophoresis, individual bands often move slightly differently from one strain to the other. Surveying strains from our stock collection, we found that nearly all the bands of a certain pair of strains differed in their mobility. Some of these chromosome length polymorphisms segregated in a 2:2 ratio, indicating that they resulted from single structural alterations (i.e. additions or deletions). One of these was mapped on the right arm of chromosome 1. Others did not segrate in a simple 2:2 ratio. That is, there were progenies which had bands not present in either parent. We suggest that these new bands are the products of recombination between homologous chromosomes having two or more structural alterations.
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    URL: http://dx.doi.org/10.1007/BF00521262
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    Comparison of Tetrahymena ARS sequence function in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe (1988)
    Springer
    Current genetics 14 (1988), S. 225-233 
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    ISSN: 1432-0983
    Keywords: ARS ; Saccharomyces cerevisiae ; Schizosaccharomyces pombe ; Tetrahymena thermophila
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated several Tetrahymena thermophila chromosomal DNA fragments which function as autonomously replicating sequences (ARS) in the heterologous Saccharomyces cerevisiae and Schizosaccharomyces pombe selection systems. The Tetrahymena ARS sequences were first isolated in S. cerevisiae and were derived from non-ribosomal micro- and macronuclear DNA. Sequence analysis of the ARS elements identified either perfect or close matches with the 11 by S. cerevisiae ARS core consensus sequence. Subcloning studies of two Tetrahymena ARS elements defined functional regions ranging in size from 50 to 300 bp. Testing of the ARS elements in S. pombe revealed that most of the T. thermophila inserts confer ARS function in both yeasts, at least in the sense of promoting a high transformation frequency to plasmids which contain them. However, the actual sequences responsible for ARS activity were not always identical in the two yeasts.
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    Construction and characterization of a haploid strain of Saccharomyces cerevisiae that completely lacks all genomic CYH2 sequences (1988)
    Springer
    Current genetics 14 (1988), S. 325-329 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Ribosomal protein genes ; CYH2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A diploid strain of the yeast Saccharomyces cerevisiae has been constructed that has one copy of the ribosomal protein gene CYH2 completely deleted and replaced with the TRP1 gene using the method of Rothstein (1983). There are only small differences in growth rate and no detectable difference in steady state level of CYH2 mRNA between the diploid that is heterozygous for the CYH2 deletion and the parent diploid with two normal copies of this gene. This suggests that the diploid must partially compensate for the loss of one CYH2 gene. Tetrad dissection shows that haploid spores lacking the CYH2 gene cannot germinate. The lethality of this deletion can be rescued by a CYH2 cDNA on a low copy vector. Haploids which lack the genomic copy of the CYH2 gene, but contain a plasmid copy of the CYH2 cDNA are able to grow normally. These CYH2 deleted yeast haploids should be useful to analyze mutationally altered CYH2 genes and genes homologous to CYH2 from other organisms without interference from a genomic copy.
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    URL: http://dx.doi.org/10.1007/BF00419989
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    The glucose- and ethanol-dependent regulation of PDC1 from Saccharomyces cerevisiae are controlled by two distinct promoter regions (1988)
    Springer
    Current genetics 14 (1988), S. 337-344 
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    ISSN: 1432-0983
    Keywords: Yeast ; Gene regulation ; Saccharomyces cerevisiae ; PDCI promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 870 by promoter fragment of the PDC1 gene that includes the carbon source dependent regulatory regions was investigated using 5′ and 3′ promoter deletions. The results indicate that glucose and ethanol regulation of PDC1 transcription are independently controlled by distinct cis-acting regions. The consensus sequence AAATCGATA may play a role in this regulation, while the sequence (ATCA)AACCT may be important in transcription initiation.
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    A colony procedure for transformation of Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 13 (1988), S. 21-23 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Transformation ; Plasmid ; Colony ; Polyethylene glycol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A rapid and simple yeast transformation procedure has been developed using colonies on agar plates. Saccharomyces cerevisiae SHY3 cells were picked up from colonies on YPD plates grown freshly or stored at 4 °C and incubated with M13RK9-T DNA at 30 °C for 1–2 h in a solution of Li+, Ca2+, Mg2+, triacetin and polyethylene glycol. About 3,500 transformants were obtained per µg of double stranded M13RK9-T DNA. Unlike the existing spheroplast techniques, single stranded M13RK9-T DNA transformed intact cells below one-hundredth frequency of the duplex form.
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    Cosmids carrying Aspergillus terreus DNA can integrate into Saccharomyces cerevisiae chromosome XII via recombination between yeast and foreign DNAs (1988)
    Springer
    Current genetics 14 (1988), S. 183-189 
    Details
    ISSN: 1432-0983
    Keywords: Aspergillus terreus clonotheque ; Saccharomyces cerevisiae ; Homologous integration ; 2 μ circledirected chromosome destabilization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A genome clonotheque consisting of 25- to 40-kb Sau3A1 fragments of Aspergillus terreus DNA was constructed in the episomal cosmid vector pES33 containing the yeastARG4 gene. From the 475 transformants of cir° yeast strain ESH-0, 23 stable Arg + transformants were independently selected. Genetic and Southern analysis of these stable transformants showed that 39% arose as a result of recombination between cloned A. terreus DNA sequences and yeast chromosome XII. The recombination events most likely occurred in the regions of homology within the rDNA clusters of A. terreus and Saccharomyces cerevisiae.
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    Generation of an ilv bradytrophic phenocopy in yeast by antisense RNA (1988)
    Springer
    Current genetics 13 (1988), S. 283-289 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Inducible antisense gene ; Acetolactate synthase ; Bradytrophic phenocopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We report for the first time on the regulation of gene expression in yeast by antisense RNA. Chimaeric genes were constructed containing the 5′ upstream and partial coding sequence of SMR1 — a sulfometuron methyl resistant allele of the ILV2 locus. Such fragments were placed 5′ to 3′ and 3′ to 5′ under control of the GAL10 promoter and CYCl terminator in a high copy YEp plasmid. Following galactose induction only transformants containing antisense RNA genes showed biological activity against SMR1 gene expression. Antisense RNA inhibited synthesis of the SMR1 gene product acetolactate synthase and thus repressed cellular growth which resulted in a bradytrophic auxotroph revertable by addition of isoleucine and valine. Antisense RNA inhibition was enhanced in galactose medium containing sulfometuron methyl and in gcn4 cells deficient for positive regulation of the ILV2 locus. This system can be used to study factors that interfere with antisense RNA function and to assign biological function to randomly cloned DNA fragments.
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    URL: http://dx.doi.org/10.1007/BF00424421
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    A yeast strain producing lys2 deletions at a high frequency after sporulation (1988)
    Springer
    Current genetics 14 (1988), S. 331-335 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Meiosis ; Deletion mutations ; Sequence dissimilarities
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A diploid yeast strain with extensive sequence dissimilarity in homologous regions near the LYS2 locus was sporulated, and spontaneous lys2 and lys5 mutant spores, selected on α-amino adipate, were analyzed. As many as 50% of the mutant spores contained a deletion in LYS2. These deletions occurred at a frequency of 5.0 × 10−7. While deletions of various sizes and endpoints were obtained, all the deletions recovered in this study included the border between homologous and non-homologous sequences located 4 kb upstream of LYS2. Large lys2 deletions that extended into an adjacent CYH2 duplication occurred at a frequency of 2.0 × 10−7, more than 1,000 times the frequency of the CYH2-LYS2 deletions found in a related haploid strain. This high frequency of CYH2-LYS2 deletions was observed only after sporulation of the diploid strain, and was dependent upon extensive sequence dissimilarity near the LYS2 locus.
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    The allosuppressor gene SAL4 encodes a protein important for maintaining translational fidelity in Saccharomyces cerevisiae (1988)
    Springer
    Current genetics 14 (1988), S. 537-543 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Allosuppressor ; Translation ; Fidelity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Allosuppressor (sal) mutations enhance the efficiency of the yeast ochre suppressor SUQ5 and define five unlinked loci, SALT-SALS. A number of sal4 mutants were isolated and found to have pleiotropic, allele;specific phenotypes, including hypersensitivity in vivo to paromomycin and other antibiotics that stimulate translational errors in yeast. To examine further the nature of the SAL4 gene product, the wild type SAL4 gene was isolated by complementation of a conditional lethal allele sal4-2, and demonstrated to be a single copy gene encoding a single 1.6 kb transcript. Restriction mapping and DNA hybridisation analysis were used to demonstrate that the SAL4 gene is identical to the previously identified omnipotent suppressor gene SUP45 (SUPT). Our results implicate the SAL4 gene product as playing a major role in maintaining translational accuracy in yeast.
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    Hyperresistance to formaldehyde of Saccharomyces cerevisiae seems not to be correlated with the formation and removal of DNA protein cross-links (1988)
    Springer
    Current genetics 13 (1988), S. 125-128 
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    ISSN: 1432-0983
    Keywords: Formaldehyde ; DNA-protein cross-links ; Repair ; Saccharomyces cerevisiae ; Hyperresistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The formation and removal of formaldehyde-mediated DNA protein cross-linking was measured by CsCI density gradient analysis in yeast strains of differing resistance to formaldehyde. Wild-type cells and transformants made hyperresistant to formaldehyde by a multi-copy vector containing the yeast SFA gene were specifically labeled in their DNA and incubated in the presence of formaldehyde. Treatment with formaldehyde lead to the formation of equal amounts of DNA protein cross-links; subsequent liquid holding of cells for 24 h resulted in the removal of nearly all DNA protein crosslinks regardless of the original formaldehyde resistance status of the strains.
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    Chloroquine, a lysosomotropic agent, inhibits zygote formation in yeast (1988)
    Springer
    Archives of microbiology 151 (1988), S. 20-25 
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    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating ; Zygote formation ; Chloroquine ; Lysosomotropic agent ; Plasma membrane ; Cell fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Haploid cells of opposite mating type of Saccharomyces cerevisiae conjugate to form zygote. During the conjugation process, the degradation or reorganization of the cell wall and the fusion of the two plasma membranes take place. Since chloroquine inhibits cellular events associated with the reorganization of the plasma membrane, the effect of the drug on conjugation was studied. Chloroquine at a concentration, at which cell growth was not retarded, inhibited zygote formation, while it did not affect other mating functions, such as sexual agglutination, production of and response to mating pheromone. Cells in a mating culture containing chloroquine formed no “prezygote” suggesting that they were not prepared for entering into fusion process. The inhibitory effect of chloroquine was reversible as cells formed zygote when they were washed after treatment with chloroquine. Zygote formation was unaffected in cells possessing chlorquine within vacuoles after incubation with the drug in complete medium (YPD) at pH 7.5, followed by washing. This suggests that chloroquine inhibits zygote formaton by adsorbing to the plasma membrane of S. cerevisiae.
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    Yeast PAPS reductase: properties and requirements of the purified enzyme (1988)
    Springer
    Archives of microbiology 150 (1988), S. 313-319 
    Details
    ISSN: 1432-072X
    Keywords: 3′-Phosphoadenylyl sulphate reductase ; Sulphite formation ; Cysteine biosynthesis ; Thioredoxin ; Saccharomyces cerevisiae ; HPLC enzyme analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The enzymatic mechanism of sulphite formation in Saccharomyces cerevisiae was investigated using a purified 3′-phosphoadenylsulphate (PAPS) reductase and thioredoxin. The functionally active protein (MR 80–85 k) is represented by a dimer which reduces 3′-phosphoadenylyl sulphate to adenosine-3′,5′-bisphosphate and free sulphite at a stoichiometry of 1:1. Reduced thioredoxin is required as cosubstrate. Examination of the reaction products showed that free anionic sulphite is formed with no evidence for “bound-sulphite(s)” as intermediate. V max of the enriched enzyme was 4–7 nmol sulphite · min-1 · mg-1 using the homologous thioredoxin from yeast. The velocity of reaction decreased to 0.4 nmol sulphite · min-1 · mg-1 when heterologous thioredoxin (from Escherichia coli) was used instead. The K m of homologous thioredoxin was 0.6 · 10-6 M, for the heterologous cosubstrate it increased to 1.4 · 10-6 M. The affinity for PAPS remained practically unaffected (K m PAPS: 19 · 10-6 M in the homologous, and 21 · 10-6 M in the heterologous system). From the kinetic data it is concluded that the enzyme followed an ordered mechanism with thioredoxin as first substrate followed by PAPS as the second. Parallel lines in the reciprocal and a common intersect in the Hanes-plots for thioredoxin were seen as indication of a ping-pong (with respect to thioredoxin) uni-bi (with respect to PAPS) mechanism.
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    Arachidonic and oleic acids stimulate zygote formation in the presence of mating pheromone in the yeast, Saccharomyces cerevisiae (1988)
    Springer
    Archives of microbiology 149 (1988), S. 507-508 
    Details
    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Mating reaction ; Zygote formation ; Mating pheromone ; Fatty acid ; Arachidonic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Effect of exogenous fatty acids on zygote formation in Saccharomyces cerevisiae was studied. Arachidonic and oleic acids considerably stimulated zygote formation, but other fatty acids tested, linoleic, linolenic, stearic and palmitic acids, did not. Pretreatment experiments with arachidonic acid showed that the stimulation of zygote formation by the fatty acid required the presence of mating pheromone.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00446752
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  • 97
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    Osmotic pressure effects and intracellular accumulation of ethanol in yeast during fermentation (1988)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1988), S. 365-372 
    Details
    ISSN: 1476-5535
    Keywords: Osmotic pressure ; Intracellular ethanol ; Yeast ; Nutrient ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The intracellular accumulation of ethanol in yeast and its potential effects on growth and fermentation have been topics of controversy for the past several years. The determination of intracellular ethanol based on the exclusion of [14C]sorbitol to estimate aqueous cell volume was used to examine the question of intracellular ethanol accumulation. An intracellular accumulation of ethanol inSaccharomyces cerevisiae was observed during the early stages of fermentation. However, as fermentation continued, the intracellular and extracellular concentrations of ethanol became similar. Increasing the osmotic pressure of the medium with glucose or sorbitol was observed to cause an increase in the intracellular ethanol concentration. Associated with this was a decrease in yeast growth and fermentation rates. In addition, increasing the osmotic pressure of the medium was observed to cause an increase in glycerol production. Supplementation of the media with excess peptone, yeast extract, magnesium sulfate and potassium phosphate was found to relieve the detrimental effects of high osmotic pressure. Under these conditions, though, no effect on the intracellular and extracellular ethanol distribution was observed. These results indicate that nutrient limitation, and not necessarily intracellular ethanol accumulation, plays a key role during yeast fermentations in media of high osmolarity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569575
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  • 98
    Electronic Resource
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    Gating behaviors of a voltage-dependent and Ca2+-activated cation channel of yeast vacuolar membrane incorporated into planar lipid bilayer (1988)
    Springer
    The journal of membrane biology 106 (1988), S. 47-55 
    Details
    ISSN: 1432-1424
    Keywords: vacuole ; lipid bilayer ; K-channel ; single channel ; DIDS ; yeast ; Saccharomyces cerevisiae ; Ca2+ activation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary A voltage-dependent and Ca2+-activated cation channel found in the vacuolar membrane of the yeast,Saccharomyces cerevisiae, was incorporated into planar lipid bilayer and its gating characteristics were studied at the macroscopic and single-channel levels. The open-channel probability at steady state, which was estimated by the macroscopic current measurement, gave a maximum value at −10 mV and decreased in a graded fashion as the voltage became more positive or more negative. The steady-state voltage dependence was explained by assuming two independent gates, which had different rate constants and opposite voltage dependence. The fast-responding gate opened when the voltage of thecis side (the side to which the vesicles were added) was made more negative and the slow-responding gate behaved in the opposite direction. Relatively high concentrations of Ca2+, about 1mm, were required on thecis side for opening the slow gate in a voltage-dependent manner. DIDS increased the open-channel probability of the fast gate when added to thecis side, but was ineffective on the slow gate.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01871766
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  • 99
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    Distributional pattern of oxytocin- and vasopressin-immunoreactivity in the neurohypophysis of the Djungarian hamster (Phodopus sungorus) (1988)
    Springer
    Cell & tissue research 253 (1988), S. 677-681 
    Details
    ISSN: 1432-0878
    Keywords: Neurohypophysis ; Hypothalamo-neurohypophyseal system ; Vasopressin ; Oxytocin ; Immunohistochemistry ; Phodopus sungorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The topography of oxytocin (OT)- and vasopressin (VP)-containing axons of the hypothalamo-neurohypophyseal system was studied in the neurohypophysis of the Djungarian hamster (Phodopus sungorus) by means of immunohistochemistry. Compared with other mammalian species, the neurohypophysis of Phodopus shows some peculiarities. Accumulations of OT-immunoreactivity around the distal vessels of the primary portal plexus can be observed in the distal median eminence and neural stem. This staining pattern indicates that OT is secreted into portal blood. In the neural lobe, OT- and VP-immunopositive fibers terminate in different areas. The vast majority of the OT-containing axons is distributed in the dorsal part of the neural lobe. In contrast, VP-containing axons are mainly found in the centre of the neural lobe up to the pars intermedia.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00219760
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  • 100
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    Immunoreactivity to vasoactive intestinal polypeptide (VIP) and thyreotropin-releasing hormone (TRH) in hypothalamic neurons of the domesticated pigeon (Columba livia). Alterations following lactation and exposure to cold (1988)
    Springer
    Cell & tissue research 251 (1988), S. 485-494 
    Details
    ISSN: 1432-0878
    Keywords: Immunohistochemistry ; Light microscopy ; Neuroendocrine regulation ; Neuropeptides ; Columba livia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of VIP- and TRH-immunoreactivity in neurons and processes within the hypothalamus of the pigeon was investigated with light-microscopic immunocytochemical techniques. Most of the VIP-containing neurons are concentrated in the middle and caudal parts of the hypothalamus, with the greatest concentration of perikarya occurring in the medial and lateral part of the ventromedial hypothalamic nucleus and the infundibular nucleus. These cells give rise to axons that seem to extend into the median eminence. An extensive network of VIP-immunoreactive fibers and varicosities occupy the external layer of the median eminence. The majority of TRH-containing neurons is found in the anterior hypothalamus with the greatest concentration of cells in the magnocellular preoptic, medial preoptic, suprachiasmatic and paraventricular nuclei. TRH-immunoreactive fibers and varicosities form a dense arborization in the external layer of the median eminence. Lactation seems to induce substantial changes in VIP as well as in TRH-immunostaining in the median eminence and other hypothalamic regions as compared to control, sexually active animals. Furthermore, TRH-immunoreactivity decreased in the median eminence following 60-min exposure to cold. These results suggest that VIP- and TRH-containing pathways in the pigeon hypothalamus are involved in the mediation of neuroendocrine responses.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00215858
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