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  • Articles  (279)
  • Rat  (197)
  • Chemical Engineering
  • Electronic structure and strongly correlated systems
  • Saccharomyces cerevisiae
  • Springer  (279)
  • 2000-2004  (25)
  • 1980-1984  (254)
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  • Articles  (279)
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  • 1
    Electronic Resource
    Electronic Resource
    A gene of the major facilitator superfamily encodes a transporter for enterobactin (Enb1p) in Saccharomyces cerevisiae (2000)
    Springer
    BioMetals 13 (2000), S. 65-72 
    Details
    ISSN: 1572-8773
    Keywords: major facilitator superfamily ; iron transport ; siderophores ; enterobactin ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract While in fungi iron transport via hydroxamate siderophores has been amply proven, iron transport via enterobactin is largely unknown. Enterobactin is a catecholate-type siderophore produced by several enterobacterial genera grown in severe iron deprivation. By using the KanMX disruption module in vector pUG6 in a fet3Δ background of Saccharomyces cerevisiae we were able to disrupt the gene YOL158c Sce of the major facilitator super family (MFS) which has been previously described as a gene encoding a membrane transporter of unknown function. Contrary to the parental strain, the disruptant was unable to utilize ferric enterobactin in growth promotion tests and in transport assays using 55Fe-enterobactin. All other siderophore transport properties remained unaffected. The results are evidence that in S. cerevisiae the YOL158c Sce gene of the major facilitator super family, now designated ENB1, encodes a transporter protein (Enb1p), which specifically recognizes and transports enterobactin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009250017785
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  • 2
    Electronic Resource
    Electronic Resource
    A polymorphism of the rat T-cell receptor β-chain variable gene 13 (BV13S1) correlates with the frequency of BV13S1-positive CD4 cells (2000)
    Springer
    Immunogenetics 51 (2000), S. 296-305 
    Details
    ISSN: 1432-1211
    Keywords: Key words Vβ13 ; CD4/CD8 ratio ; Rat ; Tcrb ; Polymorphism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Three rat BV13S1 alleles (T-cell receptor β-chain variable gene 13) were characterized by new BV13S1-allele specific monoclonal antibodies (18B1 and 17D5) and sequence analysis of expressed and genomic BV13S1. Two alleles were functional and designated BV13S1A1 present in strains LEW, BUF, PVG, and BV13S1A2 present in BN and WF. Their products differed by six amino acids, two of them in complementarity-determing region (CDR)1 and one in CDR2. A third nonfunctional allele, BV13S1A3P, was found in strains F344 and DA. Apart from a single nucleotide insertion, it was identical to BV13S1A2. All 12 rat strains tested showed association of TCRBC1 with BV8S2/4 alleles but not with the BV13S1 alleles, which may reflect a different gene order of the rat BV compared to mouse. BV13S1A1-encoded T-cell receptors (TCRs) which bind both monoclonal antibody (mAb) 18B1 and mAb 17D5 are over-represented in the CD4 lymphocyte subset. BV13S1A2-encoded TCRs which are stained by mAb 18B1 but not by mAb 17D5 show a slight CD8-biased expression. Preferential usage of BV13S1A1-positive TCRs by CD4 but not by CD8 cells in (LEW×WF)F1 hybrids and cosegregation of BV13SA1 and increased frequency of BV13S1 TCR-positive CD4 cells in a (LEW×BN)×BN backcross suggest structural differences of the two allelic products as the reason for their contrasting CD4/CD8 subset bias.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002510050623
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  • 3
    Electronic Resource
    Electronic Resource
    Localization of endothelin receptors in bleomycin-induced pulmonary fibrosis in the rat (2000)
    Springer
    Histochemistry and cell biology 122 (2000), S. 507-517 
    Details
    ISSN: 1432-119X
    Keywords: Endothelin-A receptor ; Endothelin-B receptor ; Rat ; Pulmonary fibrosis ; Immunohistochemistry ; Quantitative PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: AbstractPulmonary fibrosis is characterized by excessive extracellular matrix deposition with concomitant loss of gas exchange units, and endothelin-1 (ET-1) has been implicated in its pathogenesis. Increased levels of ET-1 from tissues and bronchoalveolar lavage have been reported in patients with pulmonary fibrosis and in animal models after intratracheal bleomycin. We characterized the cellular distribution of alveolar ET receptors by immunohistochemistry in bleomycin-induced pulmonary fibrosis in the rat and determined the regulation by bleomycin of ET receptor mRNA expression in isolated alveolar macrophages and rat lung fibroblasts. We found significant increases in the numbers of fibroblasts and macrophages at day 7 compared to day 28 and control animals. ETB receptor immunoreactivity was observed on fibroblasts and invading monocytes. Isolated fibroblasts expressed both ETA and ETB receptor mRNA, and ETA receptor mRNA was upregulated by bleomycin. Isolated resident alveolar macrophages expressed neither ETA nor ETB receptor mRNA which were also not induced by bleomycin. We conclude that, while ETB receptor stimulation of fibroblasts and monocytes recruited during bleomycin-induced lung injury exerts antagonistic effects on fibroblast collagen synthesis, the observed increase in the number of fibroblasts in vivo and upregulation of fibroblast ETA receptor mRNA by bleomycin in vitro point to a predominance of the profibrotic effects of ET receptor engagement.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s00418-004-0708-7
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  • 4
    Electronic Resource
    Electronic Resource
    A screen of yeast respiratory mutants for sensitivity against the mycotoxin citrinin identifies the vacuolar ATPase as an essential factor for the toxicity mechanism (2000)
    Springer
    Current genetics 37 (2000), S. 227-284 
    Details
    ISSN: 1432-0983
    Keywords: Key words Citrinin ; Pet mutants ; Mitochondrial biogenesis ; Vacuolar ATPase ; YKL118W disruption ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In countries with a hot climate the mycotoxin citrinin represents a serious problem in fungal food-poisoning. In humans the renal system is affected the most and the mitochondrial respiratory chain was identified as a possible sensitive target for this toxin. In addition, citrinin has an antifungal activity that also inhibits the growth of the yeast Saccharomyces cerevisiae. So far the precise mode of action and the subcellular targets for citrinin have not been identified. Therefore, we decided to use the model organism yeast for a genetic approach to identify genes that play a role in the sensitivity against this mycotoxin. A large collection of conditional respiratory deficient yeast mutants was screened for sensitivity against citrinin. One special pet-ts mutant was identified that exhibited a higher sensitivity against citrinin. The genetic system of yeast allowed the isolation of the respective wild-type gene. This yeast gene encodes the Vph2p subunit that is essential for the correct assembly of the vacuolar ATPase. Isolation of the mutated gene and gene-disruption experiments of VPH2 and the partially overlapping small YKL118W gene verified this finding. The wild-type VPH2 gene restores all defects of the mutants. In contrast to this, YKL118W gave no complementation and the null mutant showed no phenotype. Thereby the yeast vacuolar ATPase was found to be important for the toxic effect of citrinin in yeast cells. The consequences of this finding for the molecular mechanism of citrinin action and its relation to the mitochondrial respiratory chain are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002940070001
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  • 5
    Electronic Resource
    Electronic Resource
    POL32, a subunit of the Saccharomyces cerevisiae DNA polymerase δ, defines a link between DNA replication and the mutagenic bypass repair pathway (2000)
    Springer
    Current genetics 38 (2000), S. 178-187 
    Details
    ISSN: 1432-0983
    Keywords: Key wordsPOL32 ; SRS2 ; DNA repair ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pol32 is a subunit of Saccharomyces cerevisiae DNA polymerase δ required in DNA replication and repair. To gain insight into the function of Pol32 and to determine in which repair pathway POL32 may be involved, we extended the analysis of the pol32Δ mutant with respect to UV and methylation sensitivity, UV-induced mutagenesis; and we performed an epistasis analysis of UV sensitivity by combining the pol32Δ with mutations in several genes for postreplication repair (RAD6 group), nucleotide excision repair (RAD3 group) and recombinational repair (RAD52 group). These studies showed that pol32Δ is deficient in UV-induced mutagenesis and place POL32 in the error-prone RAD6/REV3 pathway. We also found that the increase in the CAN1 spontaneous forward mutation of different rad mutators relies entirely or partially on a functional POL32 gene. Moreover, in a two-hybrid screen, we observed that Pol32 interacts with Srs2, a DNA helicase required for DNA replication and mutagenesis. Simultaneous deletion of POL32 and SRS2 dramatically decreases cellular viability at 15 °C and greatly increases cellular sensitivity to hydroxyurea at the permissive temperature. Based on these findings, we propose that POL32 defines a link between the DNA polymerase and helicase activities, and plays a role in the mutagenic bypass repair pathway.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002940000149
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  • 6
    Electronic Resource
    Electronic Resource
    Endopolygalacturonase genes and enzymes from Saccharomyces cerevisiae and Kluyveromyces marxianus (2000)
    Springer
    Current genetics 38 (2000), S. 264-270 
    Details
    ISSN: 1432-0983
    Keywords: Key words Endopolygalacturonase ; Saccharomyces cerevisiae ; Kluyveromyces marxianus ; Pectinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The gene encoding endopolygalacturonase (EC 3.2.1.15) has been cloned, sequenced and expressed from three strains of Saccharomyces cerevisiae (including non-secretors) and three strains of Kluyveromyces marxianus. Both control and coding regions showed small differences within each species, one including loss of a potential glycosylation site. Two non-secreting S. cerevisiae strains (FY1679 and var. uvarum) had non-transcribed copies of functional genes. Maximum enzyme activity was achieved with the S. cerevisiae FY1679 gene in an expressing vector, with an enzyme activity of 51 μmol of reducing sugar released from polygalacturonic acid μg protein−1 min−1, the highest so far reported for a yeast.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002940000160
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  • 7
    Electronic Resource
    Electronic Resource
    Recessive mutations in SUP35 and SUP45 genes coding for translation release factors affect chromosome stability in Saccharomyces cerevisiae (2000)
    Springer
    Current genetics 37 (2000), S. 285-291 
    Details
    ISSN: 1432-0983
    Keywords: Key words Translation release factors ; Chromosome stability ; Microtubules ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chromosome stability in suppressor mutants for SUP35 and SUP45 genes coding for translation release factors was studied. We obtained spontaneous and UV-induced sup35 or sup45 mutants in a haploid strain disomic for chromosome III and tested the stability of an extra copy of this chromosome. The majority of the mutants showed increased chromosome instability. This phenotype was correlated with an increased sensitivity to the microtubule-poisoning drug benomyl which affects chromosome segregation at anaphase. Our data suggest that termination-translation factors eRF3 and eRF1 control chromosome transmission at mitotic anaphase in Saccharomyces cerevisiae.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002940050529
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  • 8
    Electronic Resource
    Electronic Resource
    Yeast Intracellular Water Determination by Thermogravimetry (2000)
    Springer
    Journal of thermal analysis and calorimetry 59 (2000), S. 643-648 
    Details
    ISSN: 1572-8943
    Keywords: drying ; intracellular water ; Saccharomyces cerevisiae ; TG
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The intracellular water content of a microorganism is an important parameter which is a determinant factor of its physiological properties. It is usually measured by complex and time consuming procedures. Thermogravimetry using infrared balance has been used for this purpose, through the identification of different drying steps occurring during the analysis. This work employs the same method with much smaller samples, using conventional thermogravimetric equipment in a simpler and faster way than other conventional procedures. Commercial yeast (Saccharomyces cerevisiae ) washed samples are analyzed in isothermal procedures which are run in about 30 min. The drying rate curve, when plotted as a function of the residual mass of the cells, allows the identification of the step where the intracellular water is lost and the determination of its content. The obtained values, on extracellular water free basis, are in the range of 65 to 69% and agree with those measured by other techniques.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1010172830355
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  • 9
    Electronic Resource
    Electronic Resource
    Molecular Modeling of the Complexes between Saccharomyces cerevisiae Phosphoenolpyruvate Carboxykinase and the ATP Analogs Pyridoxal 5′-Diphosphoadenosine and Pyridoxal 5′-Triphosphoadenosine. Specific Labeling of Lysine 290 (2000)
    Springer
    The protein journal 19 (2000), S. 67-73 
    Details
    ISSN: 1573-4943
    Keywords: Homology modeling ; rotational energy barrier ; simulated annealing ; pyridoxal 5′-diphosphoadenosine ; pyridoxal 5′-triphosphoadenosine ; Saccharomyces cerevisiae ; phosphoenolpyruvate carboxykinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Molecular mechanics calculations have been employed to obtain models of the complexes between Saccharomyces cerevisiae phosphoenolpyruvate (PEP) kinase and the ATP analogs pyridoxal 5′-diphosphoadenosine (PLP-AMP) and pyridoxal 5′-triphosphoadenosine (PLP-ADP), using the crystalline coordinates of the ATP-pyruvate-Mn2+-Mg2+ complex of Escherichia coli PEP carboxykinase [Tari et al. (1997), Nature Struct. Biol. 4, 990–994]. In these models, the preferred conformation of the pyridoxyl moiety of PLP-ADP and PLP-AMP was established through rotational barrier and simulated annealing procedures. Distances from the carbonyl-C of each analog to ε-N of active-site lysyl residues were calculated for the most stable enzyme-analog complex conformation, and it was found that the closest ε-N is that from Lys290, thus predicting Schiff base formation between the corresponding carbonyl and amino groups. This prediction was experimentally verified through chemical modification of S. cerevisiae PEP carboxykinase with PLP-ADP and PLP-AMP. The results here described demonstrate the use of molecular modeling procedures when planning chemical modification of enzyme-active sites.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007099010762
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  • 10
    Electronic Resource
    Electronic Resource
    Involvement of the Inverted Repeat of the yeast 2-micron plasmid in Flp site-specific and RAD52-dependent homologous recombination (2000)
    Springer
    Molecular genetics and genomics 263 (2000), S. 81-89 
    Details
    ISSN: 1617-4623
    Keywords: Key words Flp recombinase ; Site-specific recombination ; Homologous recombination ; RAD52 ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Site-specific recombination within the Saccharomyces cerevisiae 2-micron DNA plasmid is catalyzed by the Flp recombinase at specific Flp Recognition Target (FRT) sites, which lie near the center of two precise 599-bp Inverted Repeats (IRs). However, the role of IR DNA sequences other than the FRT itself for the function of the Flp reaction in vivo is not known. In the present work we report that recombination efficiency differs depending on whether the FRT or the entire IR serves as the substrate for Flp. We also provide evidence for the involvement of the IR in RAD52-dependent homologous recombination. In contrast, the catalysis of site-specific recombination between two FRTs does not require the function of RAD52. The efficiency of Flp site-specific recombination between two IRs cloned in the same orientation is about one hundred times higher than that obtained when only the two FRTs are present. Moreover, we demonstrate that a single IR can activate RAD52-dependent homologous recombination between two flanking DNA regions, providing new insights into the role of the IR as a substrate for recombination and a new experimental tool with which to study the molecular mechanism of homologous recombination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/PL00008678
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  • 11
    Electronic Resource
    Electronic Resource
    Ambient pH signalling in ascomycetous yeasts involves homologues of theAspergillus nidulans genes palF and palH (2000)
    Springer
    Molecular genetics and genomics 263 (2000), S. 505-513 
    Details
    ISSN: 1617-4623
    Keywords: Key wordsYarrowia lipolytica ; Saccharomyces cerevisiae ; Ambient pH signalling ; Signal transduction ; Transmembrane protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In Yarrowia lipolytica, the transcription factor Rim101p mediates both pH regulation and control of mating and sporulation. Like its homologues PacC of Aspergillus nidulans and Rim101p of Saccharomyces cerevisiae, YlRim101p is activated by proteolytic C-terminal processing, which occurs in response to a signal transduced by a pathway involving several PAL gene products. We report here the cloning and sequencing of two of these genes, PAL2 and PAL3. PAL2 encodes a putative 632-residue protein with six possible transmembrane segments, which differs from the transmembrane proteins Rim9p of S. cerevisiae and PalI of A. nidulans, but is homologous to A. nidulans PalH and to the product of the ORF YNL294c, a predicted polypeptide of unknown function in S. cerevisiae. PAL3 encodes an 881-residue polypeptide that is homologous to PalF of A. nidulans and to a newly identified putative polypeptide of S. cerevisiae. Both PAL2 and PAL3 are expressed constitutively, regardless of ambient pH. Mutations in these genes affect growth at alkaline pH and sporulation in both Y. lipolytica and in S. cerevisiae. They affect invasiveness of haploid strains in S. cerevisiae only, and conjugation in Y. lipolytica only. These results highlight the conservation of the Pal pathway initially described in A. nidulans.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004380051195
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  • 12
    Electronic Resource
    Electronic Resource
    Multiple copies of MRG19 suppress transcription of the GAL1 promoter in a GAL80 -dependent manner in Saccharomyces cerevisiae (2000)
    Springer
    Molecular genetics and genomics 262 (2000), S. 1113-1122 
    Details
    ISSN: 1617-4623
    Keywords: Key wordsGAL regulon ; Transcription ; Saccharomyces cerevisiae ; Galactose suppression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A plasmid clone that suppresses galactose toxicity in a gal7 yeast strain has been isolated from a multicopy genomic DNA library. Molecular analysis revealed that the region responsible for the suppression of galactose toxicity corresponds to the ORF YPR030w, which was named MRG19. A CEN-based plasmid carrying the above ORF was unable to suppress the toxicity. Galactokinase activity was substantially reduced in cell extracts obtained from transformants bearing multiple copies of MRG19. Multiple copies of MRG19 were also able to suppress galactokinase expression driven by the CYC1 promoter but not the TEF1 promoter. Multiple copies of MRG19 could not suppress GAL1-driven galactokinase expression in a gal80 strain. However, MRG19-mediated suppression of CYC1-driven galactokinase expression was independent of GAL80 function. These results imply that multiple copies of MRG19 suppress galactokinase expression probably at the level of transcription. In agreement with this idea, multiple copies of MRG19 also suppress β-galactosidase expression driven by the GAL1 promoter in a GAL80-dependent manner. Disruption of MRG19 leads to an increase in the cell density at stationary phase in synthetic complete medium. MRG19 encodes a previously uncharacterised 124-kDa protein that shows no sequence homology to any known proteins.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/PL00008654
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  • 13
    Electronic Resource
    Electronic Resource
    Structure-function relationships in replication origins of the yeast Saccharomyces cerevisiae: higher-order structural organization of DNA in regions flanking the ARS consensus sequence (2000)
    Springer
    Molecular genetics and genomics 263 (2000), S. 854-866 
    Details
    ISSN: 1617-4623
    Keywords: Key words Autonomously replicating sequence (ARS) ; Anti-bent DNA ; DNA structure ; Replication origin ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to better understand the involvement of the DNA molecule in the replication initiation process we have characterized the structure of the DNA at Autonomously Replicating Sequences (ARSs) in Saccharomyces cerevisiae. Using a new method for anti-bent DNA analysis, which allowed us to take into account the bending contribution of each successive base plate, we have investigated the higher-order structural organization of the DNA in the region which immediately surrounds the ARS consensus sequence (ACS). We have identified left- and right-handed anti-bent DNAs which flank this consensus sequence. The data show that this organization correlates with an active ACS. Analysis of the minimum nucleotide sequence providing ARS function to plasmids reveals an example where the critical nucleotides are restricted to the ACS and the right-handed anti-bent DNA domain, although most of the origins considered contained both left- and right-handed anti-bent DNAs. Moreover, mutational analysis shows that the right-handed form is necessary in order to sustain a specific DNA conformation which is correlated with the level of plasmid maintenance. A model for the role of these individual structural components of the yeast replication origin is presented. We discuss the possible role of the right-handed anti-bent DNA domain, in conjunction with the ACS, in the process of replication initiation, and potentialities offered by the combination of left- and right-handed structural components in origin function.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004380000253
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  • 14
    Electronic Resource
    Electronic Resource
    Characterization of staurosporine-sensitive mutants of Saccharomyces cerevisiae: vacuolar functions affect staurosporine sensitivity (2000)
    Springer
    Molecular genetics and genomics 263 (2000), S. 877-888 
    Details
    ISSN: 1617-4623
    Keywords: Key words Staurosporine ; Vacuolar-type proton pumping ATPase ; Vacuolar protein sorting ; ATP-binding cassette transporter ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mutations at several loci affect the sensitivity of the yeast Saccharomyces cerevisiae to staurosporine. We report here the characterization of novel staurosporine- and temperature-sensitive mutants (stt). Cloning and integration mapping showed that the genes STT2/STT6, STT5, STT7, STT8 and STT9 are allelic to VPS18, ERG10, GPI1, VPS34 and VPS11, respectively. The products of ERG10 and GPI1, respectively, catalyze mevalonate and glycosyl phosphatidylinositol anchor synthesis, while VPS18 and VPS11 genes belong to the class C VPS (Vacuolar Protein Sorting) genes, and the VPS34 gene is classified as a class D VPS. Therefore, staurosporine sensitivity is affected by ergosterol and glycolipid biosynthesis and by vacuolar functions. We found that other vps mutants belonging to classes C and D exhibit staurosporine sensitivity, and that they show calcium sensitivity and fail to grow on glycerol as the sole carbon source; both of the last two characteristics are shared by vacuolar H+-ATPase mutants (vma). As vma mutants were also found to show staurosporine-sensitive growth, staurosporine sensitivity is likely to be affected by acidification of the vacuole. Moreover, wild type yeast cells are more sensitive to staurosporine in alkaline media than in acidic media, suggesting that staurosporine is exported from the cytosol by H+/drug antiporters. Pleiotropic drug resistance (PDR) genes also provide some resistance to staurosporine, because Δpdr5, Δsnq2 and Δyor1 strains are more sensitive to staurosporine than the wild-type strain. This suggests that staurosporine is also exported by the ATP-binding cassette (ABC) transporters on the plasma membrane. vma mutants and vps mutants of classes C and D vps are sensitive to hygromycin B and vanadate, while ABC transporter-depleted mutants do not show such sensitivity, indicating that two systems differ in their ability to protect the cell against different types of drug.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004380000255
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  • 15
    Electronic Resource
    Electronic Resource
    MUS81 encodes a novel Helix-hairpin-Helix protein involved in the response to UV- and methylation-induced DNA damage in Saccharomyces cerevisiae (2000)
    Springer
    Molecular genetics and genomics 263 (2000), S. 812-827 
    Details
    ISSN: 1617-4623
    Keywords: Key words DNA repair ; Helix-hairpin-Helix motif ; Methylmethane sulfonate (MMS) ; Saccharomyces cerevisiae ; UV radiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The gene MUS81 (Methyl methansulfonate, UV sensitive) was identified as clone 81 in a two-hybrid screen using the Saccharomyces cerevisiae Rad54 protein as a bait. It encodes a novel protein with a predicted molecular mass of 72,316 (632 amino acids) and contains two helix-hairpin-helix motifs, which are found in many proteins involved in DNA metabolism in bacteria, yeast, and mammals. Mus81p also shares homology with motifs found in the XPF endonuclease superfamily. Deletion of MUS81 caused a recessive methyl methansulfonate- and UV-sensitive phenotype. However, mus81Δ cells were not significantly more sensitive than wild-type to γ-radiation or double-strand breaks induced by HO endonuclease. Double mutant analysis suggests that Rad54p and Mus81p act in one pathway for the repair of, or tolerance to, UV-induced DNA damage. A complex containing Mus81p and Rad54p was identified in immunoprecipitation experiments. Deletion of MUS81 virtually eliminated sporulation in one strain background and reduced sporulation and spore viability in another. Potential homologs of Mus81p have been identified in Schizosaccharomyces pombe, Caenorhabditis elegans and Arabidopsis thaliana. We hypothesize that Mus81p plays a role in the recognition and/or processing of certain types of DNA damage (caused by UV and MMS) during repair or tolerance processes involving the recombinational repair pathway.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004380000241
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  • 16
    Electronic Resource
    Electronic Resource
    Modulation of Paired-Pulse Responses in the Dentate Gyrus: Effects of Normal Maturation and Vigilance State (2000)
    Springer
    Annals of biomedical engineering 28 (2000), S. 128-134 
    Details
    ISSN: 1573-9686
    Keywords: Hippocampus ; Vigilance states ; Paired-pulse ; Dentate gyrus ; Dentate granule cells ; Evoked response ; Rat ; In vivo studies ; Perforant path ; Maturation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Abstract This study examined the effect of normal development and vigilance state on the modulation of dentate granule cell activity in the freely moving rat at 15, 30, and 90 days of age across three vigilance states: quiet waking, slow-wave sleep, and rapid eye movement sleep. Using paired-pulse stimulation, the paired-pulse index (PPI) was obtained for the dentate evoked field potentials elicited by the stimulation of the medial perforant path. Although significant differences in PPI values were observed during development, no significant vigilance state related changes were obtained. Preweaning infant rats, i.e., 15-day old, exhibited significantly less early (interpulse intervals, IPI= 20–50 ms) and late (IPI = 300–1000 ms) inhibition, and less facilitation (IPI = 50–150 ms) when compared to the 90-day old adult rats during all three vigilance states. PPI values obtained from the 30-day old group fell intermediate between the 15- and 90-day old animals. These changes in PPI values provide a quantitative measure of changes in the modulation of dentate granule cell excitability during normal maturation. They can now can be used to evaluate the impact of various insults, such as prenatal protein malnutrition or neonatal stress, on hippocampal development. © 2000 Biomedical Engineering Society. PAC00: 8717Nn, 8719La, 8719Nn
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1114/1.261
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  • 17
    Electronic Resource
    Electronic Resource
    Left Ventricular Contractility is Impaired Following Myocardial Infarction in the Pig and Rat: Assessment by the End Systolic Pressure–Volume Relation Using a Single-Beat Estimation Technique and Cine Magnetic Resonance Imaging (2000)
    Springer
    Annals of biomedical engineering 28 (2000), S. 484-494 
    Details
    ISSN: 1573-9686
    Keywords: Heart ; Left ventricle ; LV contractility ; ESPVR ; Pig ; Rat ; Magnetic resonance imaging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Abstract The end systolic pressure–volume relation (ESPVR) has been shown to be a relatively load independent measure of left ventricular (LV) contractility. Recently, several single-beat ESPVR computation methods have been developed, enabling the quantification of LV contractility without the need to alter vascular loading conditions on the heart. Using a single-beat ESPVR method, which has been validated previously in humans and assumes that normalized elastance is constant between individuals of a species, we studied the effects of myocardial infarction on LV contractility in two species, the rat and the pig. In our studies, LV pressure was acquired invasively and LV volume determined noninvasively with magnetic resonance imaging, at one week postinfarction in pigs and at 12 weeks postinfarction in rats. Normalized systolic elastance curves in both animal species were not statistically different from that of humans. Also, the slope of the ESPVR $$\left( {E_{es} } \right)$$ decreased significantly following infarction in both species, while the volume-axis intercept $$\left( {V_0 } \right)$$ was unaffected. These results indicate that a single-beat ESPVR method can be used to measure the inotropic response of the heart to myocardial infarction, and that the basis for this method (i.e., constant normalized elastance) is applicable to a variety of mammalian species. © 2000 Biomedical Engineering Society. PAC00: 8719Uv, 8761Lh, 8719Hh, 8719Rr, 8719Ff
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    URL: http://dx.doi.org/10.1114/1.289
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    Quantifying Coevolution of Nonstationary Biomedical Signals Using Time-Varying Phase Spectra (2000)
    Springer
    Annals of biomedical engineering 28 (2000), S. 1101-1115 
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    ISSN: 1573-9686
    Keywords: Time–frequency analysis ; Coherence ; Cross correlation ; Nonstationary persistent signals ; Central pattern generator ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Abstract We present a novel time-varying phase spectrum (TVPS) method to quantify the dynamics of coevolution of two persistent nonstationary coupled signals. Based on the TVPS, an instantaneous intersignal phase shift is defined within the primary frequency range in which the two signals are highly correlated. The TVPS is estimated using a fixed-window method or an adaptive-window method. In the latter method, the window length changes dynamically and automatically as a function of change in frequency of the signals. The effects of altering window types and lengths on the accuracy of the estimation of the primary phase shift is assessed by analyzing synthesized linear chirp signals with decaying amplitude and constant relative phase shift or decaying amplitude and changing relative phase shifts. The methods developed are also used for determining the evolution of the primary phase shift among ventral root activities during fictive locomotion in an in vitro rat spinal cord preparation. The analyses indicate that the TVPS method in conjunction with the determination of the primary frequency range, allows determination of both the evolution of the coupling strength and the evolution of the phase shift between two persistent nonstationary rhythmic signals in the joint time–frequency domain. An adaptive window reduces the estimation bias and the estimation variability. © 2000 Biomedical Engineering Society. PAC00: 0230-f, 8780Tq
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    URL: http://dx.doi.org/10.1114/1.1313775
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    Partial Assembly of the Yeast Mitochondrial ATP Synthase 1 (2000)
    Springer
    Journal of bioenergetics and biomembranes 32 (2000), S. 391-400 
    Details
    ISSN: 1573-6881
    Keywords: ATP synthase ; F1-ATPase ; Saccharomyces cerevisiae ; petite mutants ; epistasis ; mitochondrion ; pet mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract The mitochondrial ATP synthase is a molecular motor that drives the phosphorylation ofADP to ATP. The yeast mitochondrial ATP synthase is composed of at least 19 differentpeptides, which comprise the F1 catalytic domain, the F0 proton pore, and two stalks, oneof which is thought to act as a stator to link and hold F1 to F0, and the other as a rotor.Genetic studies using yeast Saccharomyces cerevisiae have suggested the hypothesis thatthe yeast mitochondrial ATP synthase can be assembled in the absence of 1, and even 2, ofthe polypeptides that are thought to comprise the rotor. However, the enzyme complexassembled in the absence of the rotor is thought to be uncoupled, allowing protons to freelyflow through F0 into the mitochondrial matrix. Left uncontrolled, this is a lethal process andthe cell must eliminate this leak if it is to survive. In yeast, the cell is thought to lose ordelete its mitochondrial DNA (the petite mutation) thereby eliminating the genes encodingessential components of F0. Recent biochemical studies in yeast, and prior studies in E. coli,have provided support for the assembly of a partial ATP synthase in which the ATP synthaseis no longer coupled to proton translocation.
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    URL: http://dx.doi.org/10.1023/A:1005532104617
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    Electron microscopy of the K2 killer effect of Saccharomyces cerevisiae T206 on a mesophilic wine yeast (2000)
    Springer
    Antonie van Leeuwenhoek 78 (2000), S. 117-122 
    Details
    ISSN: 1572-9699
    Keywords: electron microscopy ; killer effect ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A mesophilic wine yeast, Saccharomyces cerevisiae CSIR Y217 K − R − was subjected to the K2 killer effect of Saccharomyces cerevisiae T206 K + R + in a liquid grape medium. The lethal effect of the K2 mycoviral toxin was confirmed by methylene blue staining. Scanning electron microscopy of cells from challenge experiments revealed rippled cell surfaces, accompanied by cracks and pores, while those unaffected by the toxin, as in the control experiments, showed a smooth surface. Transmission electron microscopy revealed that the toxin damaged the cell wall structure and perturbed cytoplasmic membranes to a limited extent.
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    URL: http://dx.doi.org/10.1023/A:1026588220367
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    Pseudohyphal growth is induced in Saccharomyces cerevisiae by a combination of stress and cAMP signalling (2000)
    Springer
    Antonie van Leeuwenhoek 78 (2000), S. 187-194 
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    ISSN: 1572-9699
    Keywords: cAMP ; pseudohyphae ; Saccharomyces cerevisiae ; stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In Saccharomyces cerevisiae pseudohyphae formation may be triggered by nitrogen deprivation and is stimulated by cAMP. It was observed that even in a medium with an adequate nitrogen supply, cAMP can induce pseudohyphal growth when S. cerevisiae uses ethanol as carbon source. This led us to investigate the effects of the carbon source and of a variety of stresses on yeast morphology. Pseudohyphae formation and invasive growth were observed in a rich medium (YP) with poor carbon sources such as lactate or ethanol. External cAMP was required for the morphogenetic transition in one genetic background, but was dispensable in strain Σ1278b which has been shown to have an overactive Ras2/cAMP pathway. Pseudohyphal growth and invasiveness also took place in YPD plates when the yeast was subjected to different stresses: a mild heat-stress (37 °C), an osmotic stress (1 m NACl), or addition of compounds which affect the lipid bilayer organization of the cell membrane (aliphatic alcohols at 2%) or alter the glucan structure of the cell wall (Congo red). We conclude that pseudohyphal growth is a physiological response not only to starvation but also to a stressful environment; it appears to require the coordinate action of a MAP kinase cascade and a cAMP-dependent pathway.
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    URL: http://dx.doi.org/10.1023/A:1026594407609
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    Hexose transporters of tomato: molecular cloning, expression analysis and functional characterization (2000)
    Springer
    Plant molecular biology 44 (2000), S. 687-697 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; heterologous expression ; H+/hexose symporter ; Lycopersicon esculentum ; quantitative PCR ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A full-length (LeHT2) and two partial (LeHT1 and LeHT3) cDNA clones, encoding hexose transporters, were isolated from tomato (Lycopersicon esculentum) fruit and flower cDNA libraries. Southern blot analysis confirmed the presence of a gene family of hexose transporters in tomato consisting of at least three members. The full-length cDNA (LeHT2) encodes a protein of 523 amino acids, with a calculated molecular mass of 57.6 kDa. The predicted protein has 12 putative membrane-spanning domains and belongs to the Major Facilitator Superfamily of membrane carriers. The three clones encode polypeptides that are homologous to other plant monosaccharide transporters and contain conserved amino acid motifs characteristic of this superfamily. Expression of the three genes in different organs of tomato was investigated by quantitative PCR. LeHT1 and LeHT3 are expressed predominantly in sink tissues, with both genes showing highest expression in young fruit and root tips. LeHT2 is expressed at relatively high levels in source leaves and certain sink tissues such as flowers. LeHT2 was functionally expressed in a hexose transport-deficient mutant (RE700A) of Saccharomyces cerevisiae. LeHT2-dependent transport of glucose in RE700A exhibited properties consistent with the operation of an energy-coupled transporter and probably a H+/hexose symporter. The K m of the symporter for glucose is 45 μM.
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    URL: http://dx.doi.org/10.1023/A:1026578506625
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    c-Fos Expression by Dopaminergic Receptor Activation in Rat Hippocampal Neurons (2000)
    Springer
    Molecules and cells 10 (2000), S. 546-551 
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    ISSN: 0219-1032
    Keywords: c-Fos ; Dopamine ; D1 ; Hippocampus ; Rat ; Synaptic Plasticity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract While dopamine is likely to modulate hippocampal synaptic plasticity, there has been little information about how dopamine affects synaptic transmission in the hippocampus. The expression of IEGs including c-fos has been associated with late phase LTP in the CA1 region of the hippocampus. The induction of c-fos by dopaminergic receptor activation in the rat hippocampus was investigated by using semiquantitative RT-PCR and immuno-cytochemistry. The hippocampal slices which were not treated with dopamine showed little expression of c-fos mRNA. However, the induction of c-fos mRNA was detected as early as 5 min after dopamine treatment, peaked at 60 min, and remained elevated 5 h after treatment. Temporal profiles of increases in c-fos mRNA by R(+)-SKF-38393 (50 μM) and forskolin (50 μM) were similar to that of dopamine. An increase in [cAMP] was observed in dopamine-, SKF-, or forskolin-treated hippocampal slices. By immunocytochemical studies, control hippocampal cells showed little expression of c-Fos immunoreactivity. However, when cells were treated with dopamine, an increase in the expression of c-Fos immunoreactivity was observed after treatment for 2 h. The treatment of hippocampal neurons with R(+)-SKF38393 (50 μM) or forskolin (50 μM) also induced a significant increase in c-Fos expression. These results indicate that the dopamine D1 receptor-mediated cAMP dependant pathway is associated with the expression of c-Fos in the hippocampal neurons. These data are consistent with the possible role of endogenous dopamine on synaptic plasticity via the regulation of gene expression. Furthermore, these results imply that dopamine might control the process of memory storage in the hippocampus through gene expression.
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    URL: http://dx.doi.org/10.1007/s10059-000-0546-y
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    Substrate specificity of yeast invertase in transglycosylation reactions (2000)
    Springer
    Chemistry of natural compounds 36 (2000), S. 88-89 
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    ISSN: 1573-8388
    Keywords: Saccharomyces cerevisiae ; yeast invertase ; active enzyme
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The substrate specificity of purified yeast invertase isolated fromSaccharomyces cerevisiae in transglycosylation reactions was determined. The enzyme is specific for primary alcohols. The yeast activity is a function of the alkyl length and substrate hydrophobicity (n-butyl, isobutyl, isoamyl alcohols).
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    Detection and analysis of gastrointestinal sounds in normal and small bowel obstructed rats (2000)
    Springer
    Medical & biological engineering & computing 38 (2000), S. 42-48 
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    ISSN: 1741-0444
    Keywords: Bowel sounds ; Rat ; Motility ; Body acoustics ; Signal detection ; Signal characterisation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract This study is aimed at detecting gastrointestinal sounds (GIS) and correlating their characteristics with gastrointestinal (GI) conditions. The central hypotheses are that GIS generation depends on the motility patterns and the mechanical properties of the gut, and that changes in those result in measurable differences in GIS. An animal model which included both healthy rats and those with small bowel obstruction (SBO) was developed. The acoustic bursts, of GIS were detected by amplitude thresholding the signal envelope. Three methods of envelope estimation were proposed and evaluated. Envelope estimation using a Hilbert transform was found to produce the best results in the current application. The duration and dominant frequency of each detected GIS event was estimated and clear differences between healthy and diseased rats were discovered. In the control state, GIS events were found to consistently be of relatively short duration (3–65ms). Although the majority of events in the SBO state had similar short duration, infrequent longer events were also detected and appeared to be pathognomonic. Long duration events (〉100 ms) occurred in each of seven obstructed, but in none of 14 non-obstructed, cases (p〈0.001). It is concluded that GIS analysis may prove useful in the non-invasive, rapid, and accurate diagnosis of SBO.
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    Corticotropin releasing factor increases the adrenocortical responsiveness to adrenocorticotropin (1984)
    Springer
    Cellular and molecular life sciences 40 (1984), S. 1004-1006 
    Details
    ISSN: 1420-9071
    Keywords: Rat ; adrenocortical responsiveness ; ACTH ; plasma ; corticosterone ; plasma ; corticotropin releasing factor (CRF)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the course of studying the plasma adrenocorticotropic hormone (ACTH) and corticosterone responses to synthetic corticotropin releasing factor (CRF), we noted some disparity in the responses. A higher dose (20 μg compared with 5 μg per rat i.a.) produced an equal plasma ACTH but greater plasma corticosterone response in adult male rats. Thus, we examined the possibility that CRF increases adrenocortical responsiveness to ACTH. CRF significantly (p〈0.0005) increased the plasma corticosterone response to ACTH in rats pretreated with dexamethasone. Thus, synthetic CRF increases corticosterone secretion in rats not only by stimulating ACTH secretion, but also by increasing the adrenocortical responsiveness to ACTH.
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    Inhibition ofSaccharomyces cerevisiae division by 5-trifluoro-methyl-6-azauracil (1984)
    Springer
    Cellular and molecular life sciences 40 (1984), S. 1159-1161 
    Details
    ISSN: 1420-9071
    Keywords: Saccharomyces cerevisiae ; 5-trifluoromethyl-6-àzauracil ; yeast cell cultures ; cell division ; inhibition of
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cell division, as studied in asynchronous cultures of yeast cells, is sensitive to 5-trifluoromethyl-6-azauracil (F3CAzU). Under defined conditions (10 mmoles l−1 F3CAzU) this compound blocks immediately and completely the process of cell division. Using synchronized cells, the time-point at which division process of yeast cell can be inhibited by F3CAzU has been determined. The inhibitor effect of this compound is completely reversed by thymine, thymidine and uracil.
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    URL: http://dx.doi.org/10.1007/BF01971472
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    Do prostaglandins mediate the somatostatin preventive effect on gastric lesion? (1984)
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    Cellular and molecular life sciences 40 (1984), S. 974-975 
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    ISSN: 1420-9071
    Keywords: Rat ; prostaglandins ; gastric lesion ; intragastric distension model ; stress model ; indomethacin ; somatostatin preventive effect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The inhibition of endogenous prostaglandin synthesis by indomethacin treatment blocks the somatostatin preventive effect on the gastric lesions induced in a stress model and has no preventive effect on an intragastric distension model.
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    URL: http://dx.doi.org/10.1007/BF01946466
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    Intracerebroventricular injection of cerebrospinal fluid (CSF) from a patient with congenital indifference to pain induces analgesia in rats (1984)
    Springer
    Cellular and molecular life sciences 40 (1984), S. 1365-1366 
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    ISSN: 1420-9071
    Keywords: Rat ; cerebrospinal fluid, human ; analgesia ; naloxone ; pain indifference, congenital ; opiates, endogenous
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary CSF from a patient with congential indifference to pain was found to produce analgesia in the rat following intracerebroventricular injections. The analgesic effect was attenuated by pretreatment with naloxone suggesting the involvement of hyperactive endogenous opiate mechanisms in this patient.
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    15-Hydroxyprostaglandin dehydrogenase and hexokinase in spontaneously hypertensive rat kidney (1984)
    Springer
    Cellular and molecular life sciences 40 (1984), S. 1368-1369 
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    ISSN: 1420-9071
    Keywords: Rat ; kidney ; hypertensive ; prostaglandin dehydrogenase ; hexokinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary 15-Hydroxyprostaglandin dehydrogenase (PGDH) surged in hypertensive (SHR) and normotensive (WKY) rat kidney at 8 days of age, is greatest in SHR. Hexokinase fell in SHR at 17 days of age, but thereafter was similar to WKY. This suggests multisystem enzymatic abnormalities in SHR kidney during development of hypertension.
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    Ethanol preference in rats with a prior history of acetaldehyde self-administration (1984)
    Springer
    Cellular and molecular life sciences 40 (1984), S. 1008-1010 
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    ISSN: 1420-9071
    Keywords: Rat ; ethanol preference ; acetaldehyde self-administration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Peripherally self-injected acetaldehyde in interaction with environmental and nutritional variables significantly enhances alcohol drinking in rats and suggests an involvement of acetaldehyde in voluntary alcohol intake.
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    Cloning and integrative deletion of the RAD6 gene of Saccharomyces cerevisiae (1984)
    Springer
    Current genetics 8 (1984), S. 559-566 
    Details
    ISSN: 1432-0983
    Keywords: DNA repair ; Saccharomyces cerevisiae ; Cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Three overlapping plasmids were isolated from a YEp24 library, which restore Rad+ functions to rad6-1 and rad6-3 mutants. Different subclones were made and shown to integrate by homologous recombination at the RAD6 site on chromosome VII, thus verifying the cloned DNA segments to be the RAD6 gene and not a suppressor. The gene resides in a 1.15 kb fragment, which restores Rad+ levels of resistance to U.V., MMS and γ-rays to both rad6-1 and rad6-3 strains. It also restores sporulation ability to rad6-1 diploids. Integrative deletion of the RAD6 gene was shown not to be completely lethal to the yeast. Our results suggest that the RAD6 gene has some cell cycle-specific function(s), probably during late S phase.
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    Purification and genetic control of α-pheromone-inactivating glycoproteins in the yeast Saccharomyces cerevisiae (1984)
    Springer
    Current genetics 9 (1984), S. 39-45 
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    ISSN: 1432-0983
    Keywords: α-Pheromone-inactivating glycoproteins ; bar1-1 ; Barrier proteins ; Purification ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two kinds of a-mating-type-specific proteins inactivating α pheromone (α factor) were purified from heat shock extract of MATa cells. Their molecular weights were estimated to be 400,000 and 200,000 by gel filtration. Both proteins were detected in MATa SST1 cells but not in MATα SST1, MATa sst1-1 and MATa/MATα SST1/SST1 cells. In addition, the proteins were detected in matα2-1 SST1 cells but not in matα1-2 SST1 cells. From these results, it is concluded that these proteins are synthesized under the control of the SST1 gene and responsible for the Barrier action of MATa cells. The relationship of these proteins to the secreted Barrier protein having a higher molecular weight is discussed.
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    Structure and function of the TRP3 gene of Saccharomyces cerevisiae: Analysis of 3′- and 5′-deletions in vivo and in vitro (1984)
    Springer
    Current genetics 8 (1984), S. 173-180 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; TRP3 gene ; Deletion analysis ; Enzyme function
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two sets of deletions, entering the TRP3 gene of Saccharomyces cerevisiae from the 3′- and the 5′-end were constructed. Complementation analysis with chromosomal trp3A, trp3B and trp3C mutations was done by introducing the 3′- and 5′-truncated gene on a multicopy 2 μm-vector. The N-terminal glutamine amido transferase function is encoded by a DNA fragment of 600–700 bp, and the C-terminal indole-3-glycerol-phosphate synthase function by a DNA fragment of about 900 bp, whereas both functions together are encoded by a contiguous DNA fragment of about 1,500 bp. The bi functional TRP3-peptide thus could be dissected into two catalytically independent peptides in vivo. For the indole-3-glycerol-phosphate synthase activity, independent catalytic activity was also demonstrated in vitro: deletions entering the TRP3 gene from the 5′-end, and lacking large parts of the sequence coding for the glutamine amidotransferase function, still are able to ex press a peptide exhibiting functional indole-3-glycerol phosphate synthase activity in vitro. Deletion plasmids pME505·De1C102·2μm and DelC10·2μm exhibited shorter TRP3 transcripts according to the deleted DNA-fragments (150 and 426 by respectively) but yielded peptides of invariable Mr of 35,000 d. Transcription and translation of these peptides, which probably represent the independently folding indole-3-glycerol-phosphate synthase core are discussed.
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    Cloning of a DNA fragment from Cephalosporium acremonium which functions as an autonomous replication sequence in yeast (1984)
    Springer
    Current genetics 8 (1984), S. 155-163 
    Details
    ISSN: 1432-0983
    Keywords: Cephalosporium acremonium ; Mitochondrial DNA ; Autonomous replication sequence ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A fragment of DNA which functions as an autonomous replication sequence in yeast was cloned from Cephalosporium acremonium. Mitochondrial DNA (mtDNA) was isolated from an industrial strain of C. acremonium (08G-250-21) highly developed for the production of the antibiotic, cephalosporin C. Size, 27 kb, and restriction pattern indicated this DNA was identical to mtDNA previously isolated (Minuth et al. 1982) from an ancestral strain (ATTC 14553) which produces very low amounts of cephalosporin C. A 1.9 kb Pst1 fragment of the Cephalosporium mtDNA was inserted into a Pst1 site of the yeast integrative plasmid, Ylp5, to produce a 7.5 kb plasmid, designated pPS1. The structure of pPS1 was verified by restriction analysis and hybridization. PS1 transformed Saccharomyces cerevisiae (DBY-746) to uracil prototrophy at a frequency of 272 transformants/μg DNA. Transformation frequencies of 715 transformants/μg DNA and zero were obtained for the replicative plasmid, YRp7, and the integrative plasmid YIp5, respectively. Southern hybridization and transformation of E. coli by DNA from yeast transformed by pPS1 verified that pPS1 replicates autonomously in yeast. The uracil-independent pPS1-yeast transformants were mitotically unstable. The average retention of pPS1 after three days growth in selective and non-selective medium was 4.5% and 0.4%, respectively, compared to retentions of 4.6% and 0.5% for YRp7. The properties of pPS1 were compared to those of a related plasmid, pCP2. pCP2 was constructed (Tudzynski et al. 1982) by inserting the C. acremonium 1.9 kb Pst1 fragment into the yeast integrative plasmid, pDAM1.
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    Cloning by genetic complementation and restriction mapping of the yeast HEM1 gene coding for 5-aminolevulinate synthase (1984)
    Springer
    Current genetics 8 (1984), S. 327-331 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; 5-aminolevulinate synthase ; Cloned gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned the structural gene HEM1 for 5-aminolevulinate (ALA) synthase from Saccharomyces cerevisiae by transformation and complementation of a yeast hem1–5 mutant which was previously shown to lack ALA synthase activity (Urban-Grimal and Labbe Bois 1981) and had no immunodetectable ALA synthase protein when tested with yeast ALA synthase antiserum. The gene was selected from a recombinant cosmid pool which contained wild-type yeast genomic DNA fragments of an average size of 40 kb. The cloned gene was identified by the restauration.of growth on a non fermentable carbon source without addition of exogenous ALA. Sub cloning of partial Sau3A digests and functional analysis by transformation allowed us to isolate three independent plasmids, each carrying a 6 kb yeast DNA fragment inserted in either orientation into the single BamHI site of the vector pHCG3 and able to complement hem1–5 mutation. Analysis of the three plasmids by restriction endonucleases showed that HEM1 is contained within a 2.9 kb fragment. The three corresponding yeast trans formants present a 1, 2.5 and 16 fold increase in ALA synthase activity as compared to the wild-type strain. The gene product immunodetected in the transformant yeast cells has identical size as the wild-type yeast ALA synthase and its amount correlates well with the increase in ALA synthase activity.
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    An improved isolation procedure for yeast two-micrometer minichromosomes (1984)
    Springer
    Current genetics 9 (1984), S. 107-111 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; 2 μm minichromosomes ; Metrizamide gradients
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two micrometer minichromosomes from Saccharomyces cerevisiae were isolated without detergent using metrizamide gradients. 2 μm minichromosomes showed a lower density in metrizamide gradients relative to genomic chromatin. Our results suggest a lower ratio of proteins to DNA in 2-μm minichromosomes as compared with genomic chromatin. The procedure described herein yields minichromosomes free of cellular chromatin and ribosomal protein contamination. This method may be useful for the isolation and characterization of other yeast minichromosomes.
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    Homoplasmic yeast cells contain no selectable “hidden” mitochondrial alleles (1984)
    Springer
    Current genetics 8 (1984), S. 81-84 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Mitochondrial genes ; Vegetative segregation ; Uniparental inheritance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Zygotes of Saccharomyces cerevisiae that are heteroplasmic for mitochondrial alleles produce diploid progeny that are homoplasmic for one allele or the other, judged by the criterion that upon further subcloning they produce daughter cells of only one phenotype or the other. Here we show that when such cells are subjected to strong selection for the missing allele, it cannot be detected, so that it is probably not present in even a single copy.
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    Regulation of transcription of the Saccharomyces cerevisiae CYC1 gene: Identification of a DNA region involved in heme control (1984)
    Springer
    Current genetics 8 (1984), S. 45-48 
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    ISSN: 1432-0983
    Keywords: Iso-1-cytochrome c ; Saccharomyces cerevisiae ; Heme ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A Saccharomyces cerevisiae mutant (hem1 cycl-1) was transformed with plasmids bearing a chromosomal centromer (CEN3) and a 2 μm DNA replication origin. In one of the plasmids a functional CYC1 gene was present, in a second plasmid an XhoI fragment located between bases -245 and -678 upstream from the translation initiation codon had been deleted, in a third plasmid this region had been inverted. Results of hybridization experiments carried out with mRNA isolated from heme-deficient and heme-containing transformants indicated that heme controls transcription of the CYC1 gene and that DNA sequences located within the upstream XhoI fragment are involved in activation of the gene by heme.
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    URL: http://dx.doi.org/10.1007/BF00405431
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    Structure and function of the TRP3 gene of Saccharomyces cerevisiae: Analysis of transcription, promoter sequence, and sequence coding for a glutamine amidotransferase (1984)
    Springer
    Current genetics 8 (1984), S. 165-172 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; TRP3 gene ; Sequence analysis ; Enzyme function
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The structure and function of the TRP3 gene of Saccharomyces cerevisiae were analyzed. Subcloning of an original 4.8 kb BamHI DNA fragment, carrying the yeast TRP3 gene, allowed for a localization of the gene on a 2.5 kb ClaI/BamHI fragment. Transcription was found to proceed from the ClaI site towards the BamHI site. Three major transcription start sites were determined at positions −92, −87, and −81 by S1-mapping. The synthesis of the TRP3 gene is regulated by the general control, and was found to take place- at the transcriptional level. The sequence of the 5′-noncoding region up to position −400 and part of the coding region to position 840 were determined. The 5′-noncoding region contains sequences common to most amino acid biosynthetic genes known so far, namely a presumptive ribosome binding site, “Goldberg-Hogness boxes”, and a consensus sequence, possibly involved in the general control. For the coding region a single open reading frame was found. The deduced amino acid sequence was aligned with homologous amino acid sequences of Neurospora crassa, Pseudomonas putida and Escherichia coli. The exceptionally high homology (40–60%) between these sequences led us to postulate that the TRP3 gene product is of the structure NH2-glutamine amidotransferase-indole-3-glycerol-phosphate synthase-COOH.
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    Cloning and identification of a DNA fragment coding for the sup1 gene of Saccharomyces cerevisiae (1984)
    Springer
    Current genetics 8 (1984), S. 467-470 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Cloning ; Suppressor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A plasmid, pYsup1-1, containing a DNA fragment able to suppress the recessive mutant phenotype of the suppressor locus sup1 (allele sup1-ts36) of Saccharomyces cerevisiae was isolated from a bank of yeast chromosomal DNA cloned in cosmid p3030. The complementing gene was localized on a 2.6 kb DNA fragment by further subcloning. Evidence is presented that the cloned DNA segment codes for the sup1 structural gene (chromosome IIR).
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    Hybridization of Saccharomyces cerevisiae with Candida utilis through protoplast fusion (1984)
    Springer
    Current genetics 8 (1984), S. 575-580 
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    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Candida utilis ; Protoplast fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Auxotrophic mutants of Saccharomyces cerevisiae and Candida utilis were hybridized through protoplast fusion. Spontaneous, UV- and FPA-induced mitotic segregation indicated that after cell fusion, exclusion of the S. cerevisiae nucleus or nuclear fusion followed by preferential loss of S. cerevisiae chromosomes can take place. Some of the hybrids were stable. One of them, expressed mating and sporulation functions of the S. cerevisiae parent. Thus, markers from both parents could be recovered as mitotic and meiotic segregants.
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    Selective inhibition of transition from sexual agglutination to zygote formation by ethyl N-phenylcarbamate in Saccharomyces cerevisiae (1984)
    Springer
    Archives of microbiology 137 (1984), S. 188-193 
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    ISSN: 1432-072X
    Keywords: Agglutination substance ; α Pheromone ; Cell cycle ; Ethyl N-phenylcarbamate ; Mating reaction ; Microtubules ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Effects of ethyl N-phenylcarbamate (EPC) on the mating reaction of Saccharomyces cerevisiae were studied, with special attention on the effect on the α pheromone action. EPC inhibited zygote formation at a concentration which promoted induction of sexual agglutinability. EPC enhanced agglutinability induction by α pheromone, but inhibited α-pheromone-induced formation of large pearshaped cells in a mating type. The enhancement of agglutinability induction was accompanied with increased production of a agglutination substance and inhibition of α pheromone inactivation. EPC arrested the cell cycle of a cells probably in the step controlled by CDC19, CDC35, cAMP etc., just before the step controlled by CDC28, α pheromone etc.
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    An examination of factors affecting the instability of Saccharomyces cerevisiae glucan synthetase in cell free extracts (1984)
    Springer
    Archives of microbiology 137 (1984), S. 209-214 
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    ISSN: 1432-072X
    Keywords: Saccharomyces cerevisiae ; Glucan synthetase ; EDTA ; Magnesium ; Sucrose ; Fluoride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Yeast β(1–3) glucan synthetase is stimulated and stabilized by EDTA. Sucrose protects the enzyme from selfinactivaton. Preincubation of cell free extracts at low sucrose concentrations indicates a slow transition of the enzyme towards dissociation. Transition kinetics at 30° C and 0° C in the presence and in the absence of sucrose are interpreted assuming that a subunit is thermolabile in the free state and that sucrose increases its stability. Magnesium is deletereous for glucan synthetase in cell-free extracts. Chaotropic agents inactivate glucan synthetase according to their capacity to solubilize and depolymerize biological compounds. Fluoride plays a special role in the activation of glucan synthetase. Its action appears to be dependent on the presence of GTP (or other nucleotides). The role of all these agents on the activity and stability of the enzyme is interpreted in a unified scheme.
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    Comparison of the killer toxin of several yeasts and the purification of a toxin of type K2 (1984)
    Springer
    Archives of microbiology 137 (1984), S. 357-361 
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    ISSN: 1432-072X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Killer toxin ; Extracellular glycoprotein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A total of 13 killer toxin producing strains belonging to the genera Saccharomyces, Candida and Pichia were tested against each other and against a sensitive yeast strain. Based on the activity of the toxins 4 different toxins of Saccharomyces cerevisiae, 2 different toxins of Pichia and one toxin of Candida were recognized. The culture filtrate of Pichia and Candida showed a much smaller activity than the strains of Saccharomyces. Extracellular killer toxins of 3 types of Saccharomyces were concentrated and partially purified. The pH optimum and the isoelectric point were determined. The killer toxins of S. cerevisiae strain NCYC 738, strain 399 and strain 28 were glycoproteins and had a molecular weight of Mr=16,000. The amino acid composition of the toxin type K2 of S. cerevisiae strain 399 was determined and compared with the composition of two other toxins.
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    Isolation, and biochemical and biological characterization of an a-mating-type-specific glycoprotein responsible for sexual agglutination from the cytoplasm of a-cells, in the yeast Saccharomyces cerevisiae (1984)
    Springer
    Archives of microbiology 140 (1984), S. 113-119 
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    ISSN: 1432-072X
    Keywords: Agglutination substance ; Cell-cell recognition ; Glycoprotein ; Mating ; Saccharomyces cerevisiae ; Sexual agglutinability ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An a-mating-type-specific substance responsible for sexual agglutination was purified to 397-times in specific activity (units/mg protein) from the cytoplasm of a-mating type cells. The purified substance gave a single band stained with PAS reagent but not with both Coomassie brilliant blue and silver staining reagent by polyacrylamide gel electrophoresis in the presence of 8 M urea. However, incorporation of [35S]methionine and Lowry reaction clearly indicate that the substance is a glycoprotein. The substance specifically masked sexual agglutinability of cells of the opposite mating type α, indicating univalent action. The substance is a glycoprotein with a carbohydrate content of 90%, a pI of 4.5, and a molecular weight of 130,000. The substance was inactivated by 2-mercaptoethanol and proteolytic enzymes but not by glycolytic enzymes. The substance formed a complementary complex having no biological activity when mixed with α-agglutination substance from the wall or cytoplasm of α-cells in vitro.
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    α-mating-type-specific suppression and a-mating-type-specific enhancement by tunicamycin of sexual agglutinability in the yeast Saccharomyces cervisiae (1984)
    Springer
    Archives of microbiology 138 (1984), S. 310-314 
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    ISSN: 1432-072X
    Keywords: Glycoprotein ; Inducible strains ; Saccharomyces cerevisiae ; Sexual agglutinability ; Tunicamycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Effects of tunicamycin (TM) on the sexual agglutinability and zygote formation of Saccharomyces cerevisiae were studied using the two kinds of haploid strains, inducible and constitutive for sexual agglutinability. Induction of sexual agglutinability by opposite mating type sex pheromone of inducible strains was inhibited by TM in α mating type but not in a mating type. The recovery by temperature-shift-down from the temperature-suppressed sexual agglutinability of constitutive strains was enhanced by TM in a mating type but rather inhibited in α mating type. Pretreatment with TM of constitutive strains enhanced sexual agglutinability in a mating type but not in α mating type. The above-mentioned a-mating-type-specific agglutinability-enhancing actions of TM were discussed in relation to the action mechanism of α pheromone which induces or enhances the sexual agglutinability of a cells. Zygote formation was inhibited by TM in both constitutive and inducible strains at concentrations which showed only partially inhibitory effect on sexual agglutinability.
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    Chemical stimuli in host-habitat location byLeptopilina heterotoma (Thomson) (Hymenoptera: Eucoilidae), a parasite ofDrosophila (1984)
    Springer
    Journal of chemical ecology 10 (1984), S. 695-712 
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    ISSN: 1573-1561
    Keywords: Leptopilinaheterotoma ; Hymenoptera ; Eucoilidae ; Saccharomyces cerevisiae ; host-habitat searching ; chemoreception ; fermentation products ; ethanol ; ethyl acetate ; acetaldehyde
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Chemical stimuli play an important role in the process of searching for a host habitat by parasitic wasps. Volatile compounds originating from host habitats and/or hosts are the cues that enable such a location.Leptopilina heterotoma, a larval parasite ofDrosophila, is attracted to the food of its host, baker's yeast. Analysis of the fermentation products of baker's yeast, using a mass spectrometer, and olfactometer studies indicate that three fermentation products of this yeast, the main component of the host habitat in our laboratory, attractL. heterotoma: ethanol (5%), ethyl acetate (10−2, 10−3%), and acetaldehyde (1%). A combination of these three compounds, however, cannot compete with baker's yeast in attracting the parasites. Thus other factors, such as different compounds, concentrations, and/or combinations, also, play a role and remain to be tested.Leptopilina heterotoma does not use host-related olfactory cues in long-distance habitat location as it cannot distinguish between host habitat and host habitat with hosts.
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    Lectin-binding properties of the Merkel cell and other root sheath cells in perinatal rat vibrissae (1984)
    Springer
    Cell & tissue research 236 (1984), S. 373-381 
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    ISSN: 1432-0878
    Keywords: Merkel cell surface ; Quinacrine fluorescence ; Lectins ; Vibrissae ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Surface carbohydrates on the Merkel cell of the outer root sheath (ORS) were investigated in 1to 4-day-old rat vibrissae by use of rhodamine isothiocyanate (RITC)-conjugated lectins. The red fluorescence of RITC provided a convenient assay for lectin binding to the Merkel cell, which is itself identified by its green fluorescence following selective uptake of the dye quinacrine. In monolayers or suspensions of freshly dissociated ORS cells, the Merkel cell showed high affinity for the α-fucose-specific lectin, Ulex europeus agglutinin I (UEA-I), thus revealing a novel feature for a basally located cell. Other high-affinity lectins included concanavalin A (Con A), wheat germ agglutinin (WGA), soybean agglutinin (SBA), and Ricinus communis agglutinin I (RCA-I). In contrast, Dolichos biflorus (DBA), Bandeiraea simplicifolia I and II (BS-I and BS-II), and peanut agglutinin (PNA) virtually excluded the Merkel cell, though PNA-binding sites were unmasked after neuraminidase treatment. Other dispersed ORS cells had varying lectin affinities, and generally binding was inhibited by a competing haptenic sugar. The pattern of lectin binding seen in cryostat and paraffin sections of the vibrissa suggested that the Merkel cells share surface properties with their neighboring basal and/or spinous cells; however, unshared properties are likely to exist since ingrowing mechanosensory nerves recognize the Merkel cells, and not other epidermal cells, as their targets.
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    Administration of caerulein to rats promotes antral epithelial cell renewal (1984)
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    Cell & tissue research 236 (1984), S. 711-715 
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    ISSN: 1432-0878
    Keywords: Gastric antral mucosa ; Caerulein ; Gastrointestinal hormones ; Cholecystokinin ; Trophic effect ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The growth-promoting effect of caerulein on antral gastric mucosa was explored using Wistar rats. Implanted osmotic minipumps were used to administer submaximal doses of either caerulein or saline to normal rats for up to 4 days. In one group, reflux of bile and pancreatic juice into the stomach was avoided by previous surgical diversion of the distal common bile duct to the jejunum. DNA synthetic and mitotic activity in the antrum epithelium were estimated by 3H-thymidine pulse labelling and autoradiography during the administration of the peptide. The rate of cell migration was determined in animals killed 1, 2 and 3 days after the 3H-thymidine pulse. Administration of caerulein to normal rats provoked significant increases in both labelling and mitotic indices, and a significant acceleration of the upward cell migration in the glandular tubes. In the animals with distal diversion of bile and pancreatic secretions both labelling and mitotic indices were also increased over control values under the effect of the peptide. These data indicate that administration of caerulein stimulates cell proliferation in the antral gastric mucosa. This effect cannot be explained through increased reflux of pancreaticobiliary secretions in the stomach.
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    Morphometric studies on lipid inclusions in Sertoli cells during the spermatogenic cycle in the rat (1984)
    Springer
    Cell & tissue research 236 (1984), S. 699-709 
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    ISSN: 1432-0878
    Keywords: Testis ; Spermatogenic cycle ; Sertoli cell ; Lipid ; Morphometry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The volume and surface area of lipid inclusions often present in the cytoplasm of rat Sertoli cells was measured directly from semi-thin sections of perfusion-fixed testicular tissues using an image analyser linked to a light microscope. Sertoli cell nuclei were used as a reference for comparing any variations in the measured parameters of lipid inclusions during the rat spermatogenic cycle. Volume density of Sertoli cell lipid inclusions was assessed by morphometric analysis of Sertoli cells photographically reconstructed from electron micrographs. Maximum lipid content in Sertoli cells occurred during stages IX–XIV of the spermatogenic cycle, then declined at stages I–III and remained low from stages IV–VIII. The persistence and increase in number of many large Sertoli cell lipid inclusions beyond the stage where spermatid residual bodies are phagocytosed within the Sertoli cells (stage IX) suggests that the synthesis and lipolysis of Sertoli cell lipid inclusions represents an intrinsic functional cycle of the Sertoli cells. Stage-dependent variations in the lipid content of rat Sertoli cells offers morphological evidence that the metabolic duties of the Sertoli cells are synchronised with the spermatogenic cycle to provide local coordination of the proliferation and maturation of the germ cells.
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    The ovarian follicle as a model for the cell-mediated control of tissue growth (1984)
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    Cell & tissue research 236 (1984), S. 717-724 
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    ISSN: 1432-0878
    Keywords: Ovary ; Ovarian follicle ; Atresia ; Immunoregulation ; Immune tolerance ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Thy-1+ cells, producing Thy-1+ material, have been demonstrated by the indirect immunoperoxidase technique in the theca of growing ovarian follicles of the rat. OX-2 antigen, known as the minor glycoprotein of rat thymocytes, was detected in granulosa cells of non-growing follicles. Ia+ cells of dendritic type and/or activated macrophages were identified in the granulosa of advanced degenerating follicles, and remnants of the zona pellucida exhibited immunoglobulins. In some ovaries immunoglobulins were also bound to the zona pellucida of oocytes of early degenerating antral follicles. Medium-sized antral follicles with degenerating granulosa were occasionally invaded by cells carrying antigens of cytotoxic T lymphocytes or other T lymphocyte subsets, while degenerating large antral follicles were sometimes invaded by cells exhibiting antigen of cells with natural killer function (but not antigens of T lymphocytes). Granulosa cells of some degenerating antral follicles exhibited class-I antigens derived from the major histocompatibility complex. We suggest that cell-mediated control mechanisms of antigen expression and metabolism of tissue cells during their differentiation and degeneration should be considered in addition to the well-documented hormonal dependence of some tissues.
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    Glial cells in the pineal gland of mice and rats (1984)
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    Cell & tissue research 237 (1984), S. 245-252 
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    ISSN: 1432-0878
    Keywords: Pineal organ ; Interstitial cells ; Astrocytes ; Immunocytochemistry ; Rat ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antigenic markers characteristic of astrocytes and their differentiative states (i.e., glial fibrillary acidic protein (GFAP), vimentin, and M1 and C1 antigens) were investigated in the pineal gland of mouse and rat using double immunolabeling techniques. In both species the socalled interstitial cells as characterized by TEM were shown to be astrocytes, since they expressed vimentin, but neither fibronectin (a marker for fibroblasts and endothelial cells) nor the neuron-specific L1 antigen or tetanus toxin receptors. Subpopulations of vimentin-positive pineal astrocytes were also GFAP- and C1- antigen-positive. M1- antigenpositive cells were not detected. It is concluded that a considerable proportion of interstitial cells in the pineal gland of rat and mouse are immature astrocytes which, in contrast to other parts of the central nervous system, persist into adulthood.
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    Evidence for a role of the cytoskeleton in the in vitro folliculogenesis of the thyroid gland of the fetal rat (1984)
    Springer
    Cell & tissue research 237 (1984), S. 499-508 
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    ISSN: 1432-0878
    Keywords: Thyroid gland, fetal ; Cytoskeleton ; Cytocha lasin B ; Vinblastine ; Colchicine ; Follicular development (thyroid) ; Tissue culture ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Thyrotropic hormone (TSH) or cAMP accelerate the formation of follicular cavities in the explanted thyroid gland of the 15-day-old rat fetus. Cytochalasin B or vinblastine and nocodazole or colchicine, which disorganize microfilamental and microtubular structures respectively, inhibit or completely block in vitro-induced folliculogenesis. Exposure of the thyroid tissue to lumicolchicine, a structural isomer of colchicine deprived of antimicrotubular activity, does not inhibit the activation of folliculogenesis induced by TSH. These results are strong evidence for the supposition that microfilaments and microtubules are involved in the TSH-stimulated mechanisms resulting in thyroid folliculogenesis. Folliculogenesis requires the integrity of both microfilaments and microtubules.
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    Immunohistochemical study on the development of CRF-containing neurons in the hypothalamus of the rat (1984)
    Springer
    Cell & tissue research 238 (1984), S. 539-544 
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    ISSN: 1432-0878
    Keywords: CRF-neurons ; Hypothalamus ; Development, ontogenetic ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Appearance of immunoreactive corticotropin-releasing factor (CRF)-containing neurons was studied in developing hypothalamus of the rat by use of antisera against rat- and ovine CRF. These neurons were first recognized in the lateral and paraventricular nuclei on days 15.5 and 16.5 of gestation, respectively, when antiserum against rat CRF was employed. Antiserum against ovine CRF revealed the cells two days later exclusively in the latter nucleus. In both nuclei, the neurons increased in number with development. The neurons in the paraventricular nucleus appeared to project their immunoreactive processes to the median eminence via the periventricular and lateral pathways. In the median eminence, the immunoreaction with antiserum to rat CRF was first recognized in its anterior portion in the form of dots on day 16.5 of gestation but as beaded fibers in the external layer on day 17.5; these structures increased in amount with development in rostro-caudal direction. Although antiserum to ovine CRF was less potent in immunostainability than antiserum to rat CRF, it also revealed the beaded fibers in the median eminence on day 17.5 of gestation. Since evidence is available that the paraventricular nucleus is involved in corticotropin release, it is concluded that, in rats, the hypothalamic regulatory mechanism controlling the release of corticotropin initially appears on days 16.5–17.5 of gestation.
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    Immunocytochemical demonstration of caldesmon (a calmodulin-binding, F-actin-interacting protein) in smooth muscle fibers and absorptive epithelial cells in the small intestine of the rat (1984)
    Springer
    Cell & tissue research 235 (1984), S. 207-209 
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    ISSN: 1432-0878
    Keywords: Caldesmon ; Actin ; Immunocytochemistry ; Small intestine ; Smooth muscle ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin-interacting protein) (Sobue et al. 1982) and of actin was studied in the rat's small intestine by means of light-microscopic immunocytochemistry. Positive immunostaining for caldesmon was seen in smooth muscle cells of the intestinal wall, and of blood vessels, and in the apical portion of the absorptive epithelial cells. The immunoreactivity in goblet cells was difficult to recognize. The positive reaction to immunostaining for actin showed almost the same pattern as that for caldesmon. These results suggest that this calmodulin-binding protein may play an important role in the control of actin-myosin interaction in smooth muscle cells and in non-muscle cells.
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    Internalization of ferritin-concanavalin A by the lactating mammary cell in vivo (1984)
    Springer
    Cell & tissue research 235 (1984), S. 433-438 
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    ISSN: 1432-0878
    Keywords: Mammary gland ; Ferritin-concanavalin A ; Concanavalin A ; Endocytosis ; Membrane reuse ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ferritin-concanavalin A (Fer-Con A) was used to label the apical plasma membrane of the lactating cell to determine whether membrane internalization takes place. Rat glands were infused in vivo via the teat with 0.2 mg of Fer-Con A in 0.2 ml tris buffer (pH 7.0) containing 0.1% trypan blue, the latter acting as a marker of the infusate. Tissues were obtained from separate animals 5, 10 and 60 min postinfusion. Fer-Con A was seen in alveolar lumina bound to the outer surfaces of apical plasma membrane, microvilli and milk fat globules. It was observed within lactating cells on the inner membrane surfaces of endocytotic vesicles, Golgi cisternae, and secretory vesicles containing casein micelles, and in multivesicular bodies and lysosomes. Internalization of the ferritin-lectin conjugate into casein-containing secretory vesicles was detectable in the 5-min postinfusion tissue. Lysosomes were the only structures in control tissue that contained particles bearing some resemblance to Fer-Con A. The data provide evidence that apical plasma membrane is internalized and distributed to a number of intracellular compartments.
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    Suprachiasmatic nucleus: Numbers of synaptic appositions and various types of synapses (1984)
    Springer
    Cell & tissue research 235 (1984), S. 449-452 
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    ISSN: 1432-0878
    Keywords: Suprachiasmatic nucleus ; Morphometry ; Synapses ; Sexual dimorphism ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The suprachiasmatic nucleus (SCN) of male rats was estimated to contain 16×106 synaptic appositions (unilaterally) or 250×106 appositions in 1 mm3 tissue of the nucleus with an average of 1404 appositions per neuron. There are significantly fewer synaptic appositions in the suprachiasmatic nucleus of female rats (15×106 per SCN or 236×106 in 1 mm3 tissue of SCN with 1264 appositions per neuron on an average). Additionally, numbers of various types of synapses (axo-somatic, invaginated, dendrodendritic and optic) are estimated.
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    Regional effect of monosodium-L-glutamate on the superficial layers of superior colliculus in rat (1984)
    Springer
    Cell & tissue research 235 (1984), S. 453-457 
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    ISSN: 1432-0878
    Keywords: Monosodium-1-glutamate ; Neuropathology ; Rat ; Superior colliculus ; Toxicology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Systemic administration of monosodium-1-gluta-mate by single injections of 4 mg/g body weight in infant rats (2–10 days of age) results in acute swelling of cytoplasm and nuclear pyknosis of neurons in the stratum zonale and stratum griseum superficiale of the superior colliculus. Multiple daily doses of 4 mg/g body weight monosodium-1-glutamate result in an almost complete loss of neurons in these two superficial layers. The deeper layers appear not to be affected. No pathological effects were observed in the lateral geniculate body or pretectal complex. Light-and electron-microscopic studies reveal that the optic nerves are remarkably shrunken and many myelinated as well as unmyelinated axons are lost. Injection of 3Hproline into the vitreous body of one eye results in limited transport to the suprachiasmatic nucleus, lateral geniculate body and to lateral portions of the superior colliculus. The small percentage of intact axons in the optic nerve, as well as the limited proline transport from the eye, suggest that administration of monosodium-1-glutamate leaves intact some optic fibers, a portion of which belongs to the retinohypothalamic tract.
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    URL: http://dx.doi.org/10.1007/BF00217873
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    Electron-microscopic demonstration of the distribution of calcium deposits in the exocrine pancreas of the rat after application of carbachol, atropine, cholecystokinin, and procaine (1984)
    Springer
    Cell & tissue research 235 (1984), S. 683-690 
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    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Calcium pool ; Calcium release ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In an attempt to identify a cellular Ca2+-pool, from which calcium is released when secretagogues are applied, tissue fragments of the rat exocrine pancreas were incubated and fixed with glutaraldehyde in the presence of calcium. By means of this procedure electron-dense deposits were found on plasma membranes. X-ray microanalysis showed that these deposits contain calcium. Stimulation of tissue fragments with the use of the secretagogues carbachol or cholecystokinin reduced the number of deposits by about 80%. When the antagonist atropine was applied after carbachol stimulation, deposits reappeared on cell membranes, which then disappeared again after a second stimulation with cholecystokinin. In the presence of procaine, carbachol was inhibited and only slightly reduced the Ca2+-deposits on the plasma membranes. These results suggest that a calcium pool, from which calcium is released to induce enzyme secretion on stimulation, is located in the cell membrane
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    URL: http://dx.doi.org/10.1007/BF00226969
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    Ontogenetic appearance of somatostatin-containing nerve terminals in the median eminence of rats (1984)
    Springer
    Cell & tissue research 236 (1984), S. 47-51 
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    ISSN: 1432-0878
    Keywords: Somatostatin ; Ontogenesis ; Electron-microscopic immunohistochemistry ; Median eminence ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development of immunoreactive (ir) somatostatin-containing nerve terminals in the rat median eminence (ME) has been examined electron-microscopically. Nerve fibers containing ir particles scattered throughout the axoplasm are first seen in the external layer of the ME on day 18.5 of gestation, and, on day 21.5 appear to terminate on the basement membrane of the perivascular space of the portal vessels. After birth, the fiber terminals contain several membrane-limited granules, which are labeled with ir PAP particles. Ultrathin, Epon-embedded sections of ME, treated by the protein A gold-labeling method for somatostatin, demonstrate positively labeled granules in the nerve fibers in the postnatal ME, but in the prenatal tissue, no specific gold-labeling is found. These findings show that, in the external layer of the ME, somatostatin storing occurs in the granules in the axonal terminals after birth.
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    URL: http://dx.doi.org/10.1007/BF00216512
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    Identification of pericytes in the central nervous system by silver staining of the basal lamina (1984)
    Springer
    Cell & tissue research 236 (1984), S. 491-493 
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    ISSN: 1432-0878
    Keywords: Brain vessels ; Basal lamina ; Pericytes ; Endothelial cells ; Glial cells ; Argyrophilic staining ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Vibratome sections obtained from perfusion-fixed rat brains were stained by means of silver impregnation and physical development according to Gailyas (1970). Small pieces of the cerebral cortex were postfixed with buffered osmium tetroxide solution and processed for electron microscopy to examine the localization of the silver deposit at the cellular level. The cell surfaces of pericytes and smooth muscle cells were completely outlined by silver grains. Endothelial cells and perivascular astrocytes, however, showed an asymmetric distribution of the silver deposit, i.e., the deposit was restricted to the abluminal endothelial surface and to the astrocytic membrane adjacent to the vessel wall, respectively. The method allowed a clear-cut distinction between perikarya of endothelial cells and pericytes as well as glial cells in perivascular position, even at the light-microscopic level.
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    URL: http://dx.doi.org/10.1007/BF00214255
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    Electron-microscopic study of silver staining of nucleoli in growing oocytes of rat ovaries (1984)
    Springer
    Cell & tissue research 236 (1984), S. 249-255 
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    ISSN: 1432-0878
    Keywords: Oocyte ; Nucleolus ; Silver staining ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The nucleoli of dictyate-stage growing oocytes in rat ovaries were examined both with routine electron microscopy and electron microscopy after silver nitrate and ammoniacal silver nitrate (Ag-AS) staining. The nucleoli of the unilaminar follicular oocytes consist of twisted strands of dense fibrillar components, aggregates of granular components, and small fibrillar centers. After Ag-AS staining, silver grains are numerous on the dense fibrillar strands, fewer on the fibrillar centers, and very sporadic on the granular aggregates. The same stainability of three nucleolar components with the Ag-AS method was also confirmed in the nucleoli segregated by actinomycin D. During the transition of growing oocytes from bilaminar to plurilaminar follicle stage, the nucleolar dense fibrillar strands gradually conglomerate and are transformed into large and compact spherules. The stainability of dense fibrillar components with the Ag-AS method was lost along with this nucleolar transformation. These results may provide some new clues on the functional significance of AgAS-positive proteins in the nucleoli.
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    URL: http://dx.doi.org/10.1007/BF00214225
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    Independent development of presynaptic specializations in olfactory cortex of the fetal rat (1984)
    Springer
    Cell & tissue research 237 (1984), S. 103-109 
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    ISSN: 1432-0878
    Keywords: Synapses ; Synaptogenesis ; Development fetal ; Olfactory cortex ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Electron microscopy was used to study synaptogenesis in prepyriform cortex of fetal rat pups during early stages of synapse formation. Of special interest is the frequent occurrence of unapposed, developing synaptic specializations in axon and growth cone profiles. The location and morphology of the unapposed specializations suggests that thay are presynaptic in nature. These presumably immature presynaptic specializations are found in the lateral olfactory tract and subjacent cortex. Intermediate forms between uncontacted presynaptic specializations and definitive synapses suggest a synaptogenic sequence in which initial development of an immature presynaptic specialization begins without apposition of a postsynaptic element at that location. This implies that initiation of presynaptic development is not dependent upon postsynaptic contact and also raises the question of whether synaptic contacts could be established via presynaptic induction of postsynaptic formation.
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    URL: http://dx.doi.org/10.1007/BF00229204
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    Connexon rearrangement in cardiac gap junctions: Evidence for cytoskeletal control? (1984)
    Springer
    Cell & tissue research 237 (1984), S. 185-186 
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    ISSN: 1432-0878
    Keywords: Gap junction ; Cytoskeleton ; Heart ; Ultrarapid freezing ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using ultrarapid-freezing techniques and freezefracture electron microscopy, we report here a close association between cardiac gap junctions and specialized membrane domains containing regularly-spaced furrows. These specialized furrowed domains are observed only during periods of gap junction re-organisation (i.e., connexon redistribution) and may reflect the presence of underlying cytoskeletal elements controlling the position of connexons in the membrane.
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    URL: http://dx.doi.org/10.1007/BF00229215
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    Transendothelial channels in fenestrated endometrial blood capillaries of rats (1984)
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    Cell & tissue research 237 (1984), S. 371-373 
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    ISSN: 1432-0878
    Keywords: Endothelium ; Transport, intracellular ; Transport vesicles, channels ; Micropinocytosis ; Capillaries ; Endometrium ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Three types of transendothelial channels are described in the endothelium of blood capillaries in the endometrium of the rat. It is postulated that they may function as pores draining interstitial fluid to the venous blood.
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    Immunocytochemical demonstration of caldesmon and actin in thyroid glands of rats (1984)
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    Cell & tissue research 237 (1984), S. 375-377 
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    ISSN: 1432-0878
    Keywords: Thyroid ; Immunocytochemistry ; Caldesmon ; Actin ; Endocytosis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin interacting protein; Sobue et al. 1982) and actin was studied in the rat thyroid gland by means of light-microscopic immunocytochemistry, and the fine-structural distribution of actin filaments was examined by use of heavy meromyosin (HMM). Caldesmon and actin were demonstrated in the apical cytoplasm of almost all the follicle epithelial cells in normal as well as TSH-treated animals. Immunoreactivities for both caldesmon and actin showed almost the same pattern in localization. The smooth muscle cells of the blood vessels were also positive for caldesmon and actin. By electron microscopy, numerous actin filaments decorated by HMM and running perpendicularly or randomly to the apical surface were recognized in the apical cytoplasm of the follicle epithelial cell. These results suggest that caldesmon and actin, in conjugation with calmodulin, play a role in the regulation of cellular activity such as exocytosis and endocytosis in the apical portion of the follicle epithelial cell.
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    Effect of monensin on the Golgi apparatus of absorptive cells in the small intestine of the rat (1984)
    Springer
    Cell & tissue research 235 (1984), S. 187-194 
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    ISSN: 1432-0878
    Keywords: Golgi apparatus ; Monensin ; Small intestine ; Cytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of short-time treatment with the ionophore monensin, administered intraluminally at concentrations of 5 and 10 μM, was studied on the Golgi apparatus of absorptive cells in the small intestine of the rat. At 2–3 min after treatment most of the Golgi stacks exhibited dilated cisternae. At 4–5 min stacked cisternae were absent; they were replaced by groups of smooth-surfaced vacuoles. Dilatation and vacuolization occurred in the entire stacks without preferential effect on any particular Golgi subcompartment. Monensin did not influence the cytochemical Golgi reaction of thiamine pyrophosphatase and acid phosphatase. The characteristic staining pattern of these two enzymes in all Golgi cisternae of absorptive cells in the proximal small intestine, and the reactivity restricted to trans cisternae in distal segments of the small intestine, were unchanged after treatment with monensin. In the distal small intestine, the cytochemical pattern allowed the monensin-induced vacuoles to be attributed to the former cisor trans-Golgi face. Further, the cytochemical results demonstrate that vacuolization is not restricted to the stacked cisternae, but includes the trans-most cisterna. The latter, usually located at some distance from the Golgi stacks, has been defined as belonging to the GERL system in several types of cells. The clear response to monensin, an agent that selectively affects the Golgi apparatus, indicates common properties between trans-most and stacked Golgi cisternae.
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    Alteration by estrogen of the nucleoli in nerve cells of the rat hypothalamus (1984)
    Springer
    Cell & tissue research 235 (1984), S. 485-489 
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    ISSN: 1432-0878
    Keywords: Ventromedial nucleus ; Hypothalamus ; Ultra-structure ; Nucleoli ; Estrogen effects ; Chromatin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Estrogen is accumulated from the blood by nerve cells in the ventromedial nucleus of the hypothalamus and can facilitate female reproductive behavior by acting on this region of the brain. This cell group was examined in ovariectomized female rats, given estrogen or control treatment, by use of light and electron microscopy. A significantly greater portion of the nerve cells in the estrogen-treated animals had protuberances on their nucleolar surfaces, apparent under the light microscope. The fine structure of such protuberances included dense, aggregated material, which is shown to contain DNA by the sodium tungstate staining technique. Because increased numbers of such protuberances were found in nuclei of cells of the experimental group where previous studies demonstrated a significant increase in ultrastructural signs of biosynthetic activity, they may be associated with increased RNA synthesis. Thus, they could indicate, ultrastructurally, increased synthetic rates for RNA in nerve cells through which estrogen promotes reproductive behavior.
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    Karyometry of nuclei in liver cells (1984)
    Springer
    Cell & tissue research 235 (1984), S. 669-673 
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    ISSN: 1432-0878
    Keywords: Liver-cell heterogeneity ; Hepatic venous branches ; Karyometry ; Binucleate cells ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In untreated adult male albino rats nuclear volume and the percentage of binucleate cells were determined in the first layer of hepatocytes adjacent to hepatic venous branches of varying diameters (〈40 μm, 40 μm–80 μm, 80 μm–120μm, 120 μm–160 μm, 〉160 μm), and in the third and fourth layer of hepatocytes in the remainder of the perivenous parenchyma. In the first layer of hepatocytes adjacent to the vascular structures means of nuclear volume are significantly lower and percentage of binucleate cells significantly higher than in the cells of the remainder of the perivenous parenchyma. Within each area measured distribution curves of nuclear volume classes were homogeneous but showed heterogeneity in comparison with each other. The morphometric data presented in this study strongly support the opinion of the heterogeneity of liver cells in the perivenous zone, as previously postulated on the basis of histochemical investigations.
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    A morphometric study of the variations in subcellular structures of rat hepatocytes during 24 hours (1984)
    Springer
    Cell & tissue research 236 (1984), S. 305-315 
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    ISSN: 1432-0878
    Keywords: Hepatocytes ; Rat ; Liver ; Circadian rhythm ; Morphometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Subcellular structures of hepatocytes in periportal and perivenous zones were examined during 24 h. The volume, surface and numerical profile densities of cytoplasmic organelles were analysed morphometrically. Most subcellular structures in periportal and perivenous hepatocytes were subject to strong circadian variations. In hepatocytes from both zones, the volume densities of smooth endoplasmic reticulum (sER), mitochondria, lysosomes, peroxisomes, polysomes and lipid droplets demonstrated peak values at 16.00 h, 20.00 h or 00.00 h; trough values were at 04.00 h, 08.00 h, or 12.00 h, except for peroxisomes (16.00 h). However, the volume densities of glycogen granules and rough endoplasmic reticulum (rER) in periportal and perivenous hepatocytes exhibited maximal values at 04.00 h, 08.00 h or 12.00 h and minimal values at 20.00 h. The surface densities of sER, mitochondria, lysosomes and peroxisomes, and the numerical profile densities of mitochondria, lysosomes and peroxisomes in periportal and perivenous hepatocytes showed similar trends. These events suggest that membranes of the rER show a partial correlation with the sER, mitochondrial and lysosomal membranes during the 24-h span. This may involve the interaction between ribosomes and rER. Almost all cytoplasmic organelles examined displayed significant differences between periportal and perivenous hepatocytes, morphometrically and in fine structure, indicating that the morphofunctional variability of hepatocytes differs depending on the location in the liver acinus.
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    Mitochondrial binding of triiodothyronine (T3) (1984)
    Springer
    Cell & tissue research 236 (1984), S. 321-325 
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    ISSN: 1432-0878
    Keywords: Triiodothyronine ; Radioautography ; Mitochondria ; Liver ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To assess the distribution of the thyroid hormone triiodothyronine (T3) within intact living cells, freshly prepared dispersed rat hepatocytes were incubated with [125I]-T3 for periods of 5 min and 30 min. Lightand electron-microscopic (EM) radioautography was carried out to determine the distribution of grains over the isolated cells. Both procedures showed the grains distributed almost entirely over the cytoplasmic matrix rather than the nucleus. Grain counts under the EM were compared with expectation based on established quantitative methods. Only the mitochondria showed obvious and statistically significant grain counts, whereas the nucleus failed to accumulate grains in excess of expectations by chance alone based on area. The findings support the existence of mitochondrial binding of T3, presumably a prerequisite for its action in direct stimulation of the mitochondria.
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    Intercellular junctions between macrophages in the regional lymph node of the rat after injection of large doses of steroids (1984)
    Springer
    Cell & tissue research 236 (1984), S. 351-355 
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    ISSN: 1432-0878
    Keywords: Lymph node ; Steroids ; Macrophages ; Intercellular junctions ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Intercellular junctions were often found between macrophages in sinuses of regional lymph nodes of the rat after injection of large doses of cholesterol, cortisone acetate, and estrone at the footpad. They were identified by subplasmalemmal densities, 20–50 nm in width, beneath the plasma membranes of apposed macrophages. No distinct filamentous structures were visible in those dense regions. Electron-dense amorphous materials are lined up at the center of the intercellular space in the junctional regions. Some macrophages form clusters with intercellular junctions. No significant difference in the effect of cholesterol, cortisone acetate, and estrone on the number of intercellular junctions betwene macrophages was found.
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    Immunocytochemistry and ultrastructure of the neuropil located ventral to the rat supraoptic nucleus (1984)
    Springer
    Cell & tissue research 236 (1984), S. 171-180 
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    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Ultrastructure ; Supraoptic nucleus ; Neuropil ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The neuropil located ventral to the SON was investigated by the use of immunoperoxidase staining for neurophysins, oxytocin and vasopressin, and electron miroscopy. The study was performed in six groups of rats: 1) control; 2) infusion of isotonic saline into the CSF; 3) infusion of hypertonic saline into the CSF; 4) drinking hypertonic saline for 4 days; 5) same as group 4 but injection of colchicine into the CSF on second day of dehydration; 6) salt loading for 3 months. In the control rats the ventral neuropil contained a few immunoreactive processes, the general morphology of which was completely different from that of the neurosecretory axons emerging from the SON at its dorsal aspect. In rats of groups 3 to 6 the ventral processes (VP) became loaded with neurosecretory granules, whereas the perikarya and axons were depleted. Based on their general morphology and reactivity pattern it is suggested that the VP are dendrites. Most of these “dendrites” were embedded in a glial cushion formed by the processes of a particular type of marginal glia. Some of these “dendrites” enveloped an arteriole penetrating the optic tract. All VP were rich in synaptic contacts. The possibility that the VP of neurosecretory cells may be functionally related to the subarachnoid CSF and the arteriolar blood flow is discussed.
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    Control of peptide release from cells of the intermediate lobe of the rat pituitary (1984)
    Springer
    Cell & tissue research 236 (1984), S. 561-566 
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    ISSN: 1432-0878
    Keywords: Pituitary gland, pars intermedia ; Peptide hormones ; Dopamine ; Corticotropin-releasing hormone ; 6-Hydroxydopamine ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary When rats were injected with 6-hydroxydopamine the catecholaminergic nerve terminals in their intermediate lobes exhibited distinct signs of degeneration. Morphometric examination of the Golgi apparatus in cells of the intermediate lobe of these rats showed significant enlargement of Golgi cisternae. The release of adrenocorticotropin, β-endorphin/lipotropin and α-melanotropin from intermediate-lobe cells in vitro was measured by radioimmunoassay. The high basal peptide release was inhibited by dopamine and stimulated by methyl-isobutyl-xanthine. In contrast, γ-aminobutyric acid, serotonin, histamine and noradrenaline, or corticotropin-releasing hormone, rat hypothalamic extract and vasopressin had no or only very weak effects. These observations indicate that the synthetic apparatus of intermediate-lobe cells is constantly depressed by dopaminergic nerves. We were not able to stimulate peptide release from intermediate-lobe cells by use of the abovementioned endogenous agents.
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    Effects of prolonged maternal fast on the pancreas of 18–21-day-old foetal rats (1984)
    Springer
    Cell & tissue research 237 (1984), S. 169-179 
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    ISSN: 1432-0878
    Keywords: Foetal pancreas ; β Cells ; Insulin ; Fasting mothers ; Morphometry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary After maternal fasting for 72 h the pancreatic β cells of 18-day-old foetal rats show a conspicuous enrichment in secretory material, with an increase of pancreatic insulin concentration and a marked development of the rough endoplasmic reticulum and the Golgi apparatus. The morphometric analysis shows that the intracytoplasmic migration of the secretory granules is inhibited, principally inside the cell web. Consequently the number of secretory granules fused with plasma membrane decreases and this is associated with a decreased foetal plasma insulin. The difference in the ultrastructural aspect of the β cells of foetuses from fasting mothers and of foetuses from fed mothers is less conspicuous at 19 days of gestation and progressively disappears at 20 and 21 days. The modifications in ultrastructural aspect and in functional state are discussed.
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    Histological study of ibotenic acid-induced modifications of rat retina and their attenuation by diazepam (1984)
    Springer
    Cell & tissue research 238 (1984), S. 81-85 
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    ISSN: 1432-0878
    Keywords: Retina ; Ibotenic acid ; Toxicity ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary After 2 h intraocular injections of 19 and 190 nmoles ibotenic acid in the rat retina produced an intensive vacuolization of the inner plexiform layer and cellular alterations, in the inner nuclear layer and ganglion cell layer. These alterations consisted of either cytoplasmic swelling accompanied by clumping of the nuclear chromatin or darkening of the cytoplasm along with nuclear condensation. A week later the retinas were thinner than the controls due to the disappearance of the affected cells. Pre-treatment with diazepam prevents the morphological alterations induced by 19 nmoles ibotenic acid; mainly the swelling, which was completely prevented, while the darkening was reduced drastically, although some vacuolization of the inner plexiform layer is still present.
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    The effect of an LH pulse on 3H-thymidine incorporation into cultured ovaries of metestrous rats (1984)
    Springer
    Cell & tissue research 238 (1984), S. 159-163 
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    ISSN: 1432-0878
    Keywords: Ovary ; Rat ; Cell division ; Luteinizing hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of an LH pulse on the rate at which 3H-thymidine is incorporated into cultured ovaries of metestrous rats was studied. In comparison to ovaries cultured with tonic LH, an LH pulse (1) “rescued” follicles from atresia, (2) induced thecal cell proliferation, and (3) increased the rate at which granulosa cells enter mitosis. It is concluded that LH pulses increase follicular growth by first triggering thecal cell proliferation and then inducing mitotic divisions within the granulosa cells of both atretic and non-atretic follicles.
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    Electron-microscopic immunohistochemical study of the localization of immunoglobulin G in the choroid plexus of the rat (1984)
    Springer
    Cell & tissue research 238 (1984), S. 177-182 
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    ISSN: 1432-0878
    Keywords: Choroid plexus ; Immunoglobulin G ; Permeability ; Anti-HRP-IgG ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of autologous antiperoxidase immunoglobulin G (IgG) was studied in the choroid plexus of Lewis rats immunized against horseradish peroxidase (HRP). This experiment was performed to study the permeability of the choroid plexus to intravascular IgG. It was shown that autologous IgG was present in the extravascular spaces. The transendothelial transfer appeared to occur mainly via the fenestrations and some interendothelial junctions. No transfer of IgG at the level of epithelial cells toward the cerebrospinal fluid was demonstrated. Interstitial spaces in contact with the connective-tissue cells of the choroid stroma were strongly labeled. The significance of these spaces remains hypothetical and raises the question of the fate of IgG from the interstitial space.
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    URL: http://dx.doi.org/10.1007/BF00215159
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    Glial fibrillary acidic protein and differentiation of neonatal rat pituicytes in vitro (1984)
    Springer
    Cell & tissue research 238 (1984), S. 191-195 
    Details
    ISSN: 1432-0878
    Keywords: Glial fibrillary acidic protein (GFAP) ; Pituicytes, neonatal ; Development, ontogenetic ; Immunofluorescence ; Organ culture ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The appearance and intracellular localisation of glial fibrillary acidic protein (GFAP) in pituicytes in neural lobe cultures of newborn rats aged 7 to 30 days were investigated by use of the indirect immunofluorescence method. GFAP-immunoreactive cells were observed mostly in the outgrowth zone. GFAP was localised in the perikaryal cytoplasm as well as in pituicyte processes. GFAP-positive pituicytes showed considerable morphological polymorphism. The presence of GFAP — astrocytic marker — in pituicytes in vitro and the evident morphological similarity to cultured astrocytes suggest the astroglial character of these cells.
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    URL: http://dx.doi.org/10.1007/BF00215161
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    Localization and function of cyclic guanosine monophosphate-phosphodiesterase activity in the retinal rods of the rat by means of a newly developed cytochemical method (1984)
    Springer
    Cell & tissue research 238 (1984), S. 453-457 
    Details
    ISSN: 1432-0878
    Keywords: Cyclic guanosine monophosphate-phosphodiesterase (cGMP-PDEase) ; New cytochemical method ; Retina ; Rods, outer segments ; Light perception ; 5′GMPase ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cyclic guanosine monophosphate-phosphodiesterase (cGMP-PDEase) activity was studied histo- and cytochemically in the retinal rods of the rat with the use of a newly developed technique. Intense activity of cGMP-PDEase was evenly distributed over the outer segments of the rods. Reaction product was observed on the plasmalemma and on the disk membranes of the outer segments. A weak reaction product occurred also on the plasmalemma of the inner segments; however, no precipitate was found in the perinuclear and synaptic portions of the rod cells. The enzyme activity was strongly inhibited by 2 mM theophilline and by 2 mM 3-isobutyl-1-methylxanthine (IBMX). To confirm the specificity of this new cGMP-PDEase method, the localization of 5′nucleotidase (5′GMPase) was also studied. In contrast to the activity of cGMP-PDEase, the activity of 5′GMPase was distributed on the plasma membrane of the photoreceptor cells extending over a wide range from the synaptic endings in the outer plexiform layer to the tip of the outer segments.
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    Immunohistochemical study of S-100 protein in the postnatal development of Müller cells and astrocytes in the rat retina (1984)
    Springer
    Cell & tissue research 238 (1984), S. 503-508 
    Details
    ISSN: 1432-0878
    Keywords: S-100 ; Müller cell ; Astrocyte ; Development ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The rat retina was studied by immunohistochemistry with antibody to S-100 protein during the first three postnatal weeks. Immunoreactive astrocytes are first detected subjacent to the inner limiting membrane close to the optic disc. They gradually increase in number and spread toward the ora serrata along the inner surface of the retina as the development proceeds. S-100-immunostained Müller cells are first identified on the 12th postnatal day although their immunoreactivity is much weaker than that of astrocytes at the same stage. This differential intensity of the immunoreactivity of the two cell types facilitates observation of the entire shape of the astrocyte. This characteristic reveals that cellular investments of blood vessels in the inner retina are formed by astrocytic processes whereas those in the outer plexiform layer are derived from processes of Müller cells. The cellular investment becomes complete by the 18th postnatal day.
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    URL: http://dx.doi.org/10.1007/BF00219865
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    Effects of ovariectomy and estradiol administration on the adrenal macrophage system of the rat (1984)
    Springer
    Cell & tissue research 238 (1984), S. 559-564 
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    ISSN: 1432-0878
    Keywords: Adrenal cortex ; Macrophages ; Ovariectomy ; Rat ; Estradiol administration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Macrophages of the adrenal cortex were studied in normal male and female, ovariectomized and estradiol-injected rats. In normal male rats few macrophages with numerous granules were observed in the zona fasciculatazona reticularis border, and in the zona reticularis. Granules, identified as lysosomes, were limited by a single membrane with a heterogeneous matrix; they exhibited acid phosphatase- and aminotriazole-resistant peroxidatic activities. A larger number of macrophages had identical distributions in normal female rats. In ovariectomized and estradiol-injected rats the number and distribution of adrenal macrophages were similar to those in normal females; however, in spayed animals the number of these cells in the zona reticularis was higher than in the other experimental groups. Lysosomes in macrophages of treated animals were more numerous and their contents more complex than in normal male animals. These results indicate that the adrenal macrophage system is stimulated in experimental conditions involving high levels of circulating estrogens.
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    Fine structure of the small, granule-containing cells in the superior cervical ganglia of hydrocortisone-treated early postnatal and adult rats (1984)
    Springer
    Cell & tissue research 238 (1984), S. 297-305 
    Details
    ISSN: 1432-0878
    Keywords: Sympathetic ganglion ; Development, ontogenetic ; Corticosteroid treatment ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Hydrocortisone injections into rats on postnatal days 3–9 caused an increase in the number of small granulecontaining cells in the superior cervical ganglia. These cells, corresponding to the small, intensely fluorescent cells, showed an extensive rough endoplasmic reticulum, a large Golgi apparatus and a very large number of granular vesicles. In addition to the granular vesicles, 70–160 nm in diameter, in which the dense core filled most of the vesicle, most cells of the hydrocortisone-injected rats contained also larger granular vesicles, up to 350 nm in diameter, in which the dense core was eccentrically located. A minority of the cells contained only granular vesicles 70–100 nm in diameter, which was the only type seen in the saline-treated control rats. Thirty days after discontinuation of the hydrocortisone treatment, most of the cells with large granular vesicles had disappeared, and only two profiles of such cells were seen on day 40. The other small cells contained only granular vesicles 70–160 nm in diameter, and these cells could not be distinguished from the small granule-containing cells of 40-day-old control rats treated early postnatally with saline. Hydrocortisone treatment, first on days 3–9 and subsequently on days 40–46, caused reappearance of the small granule-containing cells with large granular vesicles up to 350 nm in diameter, the dense core of which was eccentrically located. Hydrocortisone treatment on days 40–46 only was not followed by appearance of such cells in rats treated with saline on days 3–9.
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    Compartments and perivascular arrangement of the meninges covering the cerebral cortex of the rat (1984)
    Springer
    Cell & tissue research 238 (1984), S. 459-474 
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    ISSN: 1432-0878
    Keywords: Meningeal compartment ; Perivascular space ; Brain intercellular compartment ; Cerebral cortex ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The intercellular clefts of the brain and the leptomeninges, and the perivascular spaces were studied with reference to the results obtained in a previous study (Krisch et al. 1983). The spatial relationships of these compartments were analyzed at the electron-microscopic level. Horse-radish peroxidase (HRP) was injected into the brain or into the contralateral ventricle. The pattern of distribution of HRP depends on the boundary situation in the individual compartments. The inner and outer pial layers accompany the vessels intruding into the brain. In the Virchow-Robin space the pial funnel obliterates within a short distance. The inner arachnoid layer is continuous with the outer arachnoid layer when it covers the vessels traversing the meningeal space. The perivascular compartment is not in communication with the arachnoid space; moreover, the pial funnel within the Virchow-Robin space is sealed off against the arachnoid space. Thus, blood vessels traversing the meningeal spaces and subsequently penetrating the brain surface are exposed to the common intercellular compartment represented by the intercellular clefts of the brain and the leptomeninges; this compartment does not communicate with the other compartments. The cerebrospinal fluid located in this intercellular compartment is preferentially drained into the upper cervical lymph nodes.
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    Peroxidase-positive endothelial cells in rat liver (1984)
    Springer
    Cell & tissue research 238 (1984), S. 635-642 
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    ISSN: 1432-0878
    Keywords: Liver ; Endothelium ; Kupffer cells ; Peroxidase ; Cytochemistry ; Ultrastructure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rat liver fixed by perfusion with low glutaraldehyde concentrations was incubated in diaminobenzidine-containing medium to stain for peroxidase. Endogenous peroxidatic activity was found not only in Kupffer cells but also in the endothelial cells lining the sinusoids and central veins. The reaction product was localized in the nuclear envelope and endoplasmic reticulum. The peroxidatic activity in endothelial cells showed a concentration-dependent sensitivity to glutaraldehyde: in liver samples fixed with 0.25% glutaraldehyde, approx. 23% of the sinusoidal endothelial cells and 65% of central vein endothelium were peroxidase-positive; with 0.5% glutaraldehyde, only approx. 8% of the sinusoidal endothelial cells contained detectable amounts of the reaction product; with 1.5% glutaraldehyde all endothelial cells were consistently peroxidase-negative. No peroxidatic activity could be found in liver endothelial cells following isolation by centrifugal elutriation. Endothelial cell peroxidase may possibly be involved in defense responses of liver and/or, as a part of prostaglandin synthase system, in prostanoid production.
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    Biogenesis of mitochondria: Defective yeast H+-ATPase assembled in the absence of mitochondrial protein synthesis is membrane associated (1984)
    Springer
    Journal of bioenergetics and biomembranes 16 (1984), S. 561-581 
    Details
    ISSN: 1573-6881
    Keywords: H+-ATPase complex ; assembly defect ; Saccharomyces cerevisiae ; mitochondrial biogenesis ; membrane association
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract We have investigated the extent to which the assembly of the cytoplasmically synthesized subunits of the H+-ATPase can proceed in a mtDNA-less (rho°) strain of yeast, which is not capable of mitochondrial protein synthesis. Three of the membrane sector proteins of the yeast H+-ATPase are synthesized in the mitochondria, and it is important to determine whether the presence of these subunits is essential for the assembly of the imported subunits to the inner mitochondrial membrane. A monoclonal antibody against the cytoplasmically synthesized β-subunit of the H+-ATPase was used to immunoprecipitate the assembled subunits of the enzyme complex. Our results indicate that the imported subunits of the H+-ATPase can be assembled in this mutant, into a defective complex which could be shown to be associated with the mitochondrial membrane by the analysis of the Arrhenius kinetics of the mutant mitochondrial ATPase activity.
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    Structure-activity relationship of stress-inducing odorants in the rat (1984)
    Springer
    Journal of chemical ecology 10 (1984), S. 1007-1018 
    Details
    ISSN: 1573-1561
    Keywords: Rat ; Rattus norvegicus sp. ; odorants ; stress ; behavior ; open field ; corticosterone ; fox dropping ; ketone ; sulfur ; compounds ; tans ; mercaptoketones ; repellent ; structure-activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The stress for 12 sulfur-containing synthetized volatiles was evaluated in male Wistar rats and compared to that for fox-dropping extract concentrate. Stress behavior was analyzed by quantifying various stress responses in a standard open field and measuring the increase in plasma corticosterone concentration. Nine compounds induced stress—a dihydrothiazole, two cyclic polysulfides, five mercaptoketones, and a mercaptan. For the mercaptoketones, the following structure-activity relationships were observed. Size can vary considerably; the mercapto group can be either alpha or beta and either secondary or tertiary. The keto group is not essential, since a structurally related mercaptan remains active. The mercapto group is essential for activity in mercaptoketones, since conversion to a methyl sulfide resulted in a neutral response. This type of odorant could function as an allomone and may have potential in rat control as an area repellent.
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    Der Einfluß von Cadmium, Zink, Blei und Quecksilber auf die Enzymaktivität beiSaccharomyces cerevisiae in vitro (1983)
    Springer
    European journal of nutrition 22 (1983), S. 205-212 
    Details
    ISSN: 1436-6215
    Keywords: Schwermetallwirkung ; Malatdehydrogenase ; Glutamatdehydrogenase ; Glycerinaldehyd-3-phosphatdehydrogenase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Summary The difference between cadmium, zinc, lead, and mercury in regard of their effects on the activity of the enzymes tested is very slight. Concentrations higher than 10−5 M reduce significantly the activity of the enzymes, and concentrations of approximately 10−3 M inhibit it completely. An increase of the activity cannot be detected. The addition of combinations of cadmium, zinc, and lead results in a summing up of the toxic effects, whereas the interaction between mercury and the other three heavy metals shows a cumulative effect, which is appointed nearly completely by the heavy metal more toxic. The findings suggest that under in-vitro conditions there exists a direct interaction between the heavy metals and the enzymes.
    Notes: Zusammenfassung Die vier Schwermetalle Cadmium, Zink, Blei und Quecksilber unterscheiden sich in ihrer Wirkung auf die Aktivität der untersuchten Enzyme nur sehr wenig. Konzentrationen über 10−5 M vermindern die Enzymaktivität signifikant, und Konzentrationen von etwa 10−3 M unterbinden sie völlig. Eine Steigerung der Enzymaktivität läßt sich nicht feststellen. Die Zugabe von Cadmium-, Zink- und Bleikombinationen führt zu einer Addition der toxischen Effekte, während bei der Interaktion zwischen Quecksilber und den anderen drei Schwermetallen die Gesamtwirkung fast ausschließlich durch das stärker hemmende Schwermetall allein bestimmt wird. Die erhaltenen Ergebnisse lassen vermuten, daß es unter Invitro-Bedingungen zu einer direkten Wechselwirkung zwischen den Schwermetallen und den Enzymen kommt.
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    Oral 1,25(OH)2D3: An effective prophylactic treatment for glucocorticoid osteopenia in rats (1983)
    Springer
    Calcified tissue international 35 (1983), S. 107-110 
    Details
    ISSN: 1432-0827
    Keywords: Calcium ; Glucocorticoid ; Vitamin D ; Osteoporosis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Eighty-eight adult male Sprague-Dawley rats were given a diet with either (a) 0.5% Ca and 0.6% P or (b) 0.01% Ca and 0.6% P. Osteopenia was created by adding prednisolone to the diet. The prophylactic effect of oral 1,25(OH)2D3 on the osteopenia was studied. It was found that prednisolone osteopenia in the rat was associated with defective Ca absorption. By giving an oral dose of 1,25(OH)2D3, it was possible to maintain normal Ca absorption during prednisolone treatment and to prevent the bone loss. No significant hypercalcemia or any kidney calcifications were seen. These results are in contrast to earlier findings, in which subcutaneous administration of 1,25(OH)2D3 failed to prevent prednisolone osteopenia because of its tendency to increase bone resorption.
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    Nuclear association in yeast of a hybrid vector containing mitochondrial DNA (1983)
    Springer
    Current genetics 7 (1983), S. 165-166 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Cephalosporium acremonium ; Mitochondrial hybrid vector ; Nuclear association
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The hybrid vector pCP2, consisting of the bacterial plasmid pBR325, the nuclear gene Leu-2 of Saccharomyces cerevisiae and a fragment of mitochondrial DNA from Cephalosporium acremonium, was found to associate with the nucleus in a transformed strain of Saccharomyces cerevisiae. This was inducted by (1) efficient expression of the Leu-2 gene as evidenced by a short generation time on selective medium; (2) independence of Leu-2 gene expression from mitochondrial protein synthesis, since pCP2 was shown to replicate and to be expressed in petite mutants; (3) association of pCP2 with isolated DNA from nuclei as proved by transformation experiments with E. coli.
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    Mating factor dependence of G1 cell cycle mutants of Saccharomyces cerevisiae (1983)
    Springer
    Current genetics 7 (1983), S. 309-312 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; G1 cdc mutants ; tα-factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutants in four G1 cdc strains of Saccharomyces cerevisiae were isolated which failed to show division arrest in the presence of α-factor. The cell cycle properties, terminal arrest morphology and mating competence of these mutants at the restrictive temperature were examined. The G1 specific arrest of the cdc 36 and cdc39 mutants is dependent upon the availability of an intact mating factor response system in Mat a cells. Cdc28 and cdc37 mutants exert their cell cycle blocks independently of the mating factor pathway. It is likely that the nature of the primary growth defect in cdc36 and cdc39 mutants is such that the α-factor pathway is activated in the absence of the pheromone at the restrictive temperature and that G1 arrest is a secondary consequence of a non-cycle specific event in such mutants.
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    Mitotic segregation of 2 μm-pbr322 chimaeric plasmids in yeast (1983)
    Springer
    Current genetics 7 (1983), S. 235-237 
    Details
    ISSN: 1432-0983
    Keywords: DNA replication ; Shuttle vectors ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitotic segregation of three 2 μm-pBR322 chimaeric plasmids (YEp6, YEp21, and YEp24) was studied in yeast. Each displayed a characteristic rate of loss: YEp6 was lost at approximately twice the rate of YEp21 and YEp24. The loss rates were not significantly increased when two chimaeric plasmids were coresident, nor was the endogenous 2 μm plasmid itself displaced. Therefore these plasmids appear to be compatible in yeast.
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    Analysis of the effect of radiation repair mutations on the DEL1 mutator region of Saccharomyces cerevisiae (1983)
    Springer
    Current genetics 7 (1983), S. 57-61 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; DEL1 ; rad ; ste7
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In DEL1 strains of the yeast, Saccharomyces cerevisiae, the iso-1-cytochrome c (CYC1) region is flanked on either side by Tyl elements in direct orientation which promote cyc1 deletions of the bracketed DNA in the haploid cell. In this study, we asked which genes might control this event by testing the possibility that the DEL1 mutation mechanism requires an enzyme (or enzymes) that is also utilized in the repair of damaged DNA. To this end, we independently coupled eight repair mutations, rad3–2, rad4–4, rad6–1, rad6–3, rad9–1, rev3–1, rad50–1, and rad51-1, toDEL1 and asked whether DEL1 was still functional. We found that none of these rad mutations significantly affects the mutation frequency of 10−6-10−5 established in DEL1 strains for the CYC1 locus. Furthermore, we determined that ste7, a temperature-sensitive sterile allele known to alter gene regulation in Ty-mediated mutations, is not required for DEL1 function. Finally, DEL1 is not temperature-sensitive at 23° or 37 °C.
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    Leucine biosynthesis in yeast (1983)
    Springer
    Current genetics 7 (1983), S. 369-377 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Isoenzymes ; Induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Tetrad analysis indicates that α-isopropylmalate synthase activity of yeast is determined by two separate genes, designated LEU4 and LEU5. LEU4 is identified as a structural gene. LEU5 either encodes another α-isopropylmalate synthase activity by itself or provides some function needed for the expression of a second structural gene. The properties of mutants affecting the biosynthesis of leucine and its regulation suggest that the expression of LEU1 and LEU2 (structural genes encoding isopropylmalate isomerase and β-isopropylmalate dehydrogenase, respectively) is controlled by a complex of a-isopropylmalate and a regulatory element (the LEU3 gene product). Similarities and differences between yeast and Neurospora crassa with respect to leucine biosynthesis are discussed.
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    Trehalose: Its role in germination of Saccharomyces cerevisiae (1983)
    Springer
    Current genetics 7 (1983), S. 393-397 
    Details
    ISSN: 1432-0983
    Keywords: Trehalose ; Glycogen ; Sporulation ; Germination ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mutants with specific lesions were used to differentiate between the functions of glycogen and trehalose in S. cerevisiae. Diploids which harbor the glc1/glc1 mutation depend upon the phosphorylated, less active form of glycogen synthase and show a more active, phosphorylated form, of the enzyme trehalase. These conditions are due to a lesion in the regulating subunit of the cAMP-dependent protein kinase. Such cells are unable to sporulate. Diploids which contain the sst1/sst1 mutation have normal glycogen metabolism but their trehalose-6-phosphate synthase is not active. Such strains sporulate but germination is poor and only one-spore tetrads are formed. These results confirm that glycogen is needed to trigger sporulation while trehalose plays a role in the germination process. Different systems, I and II, of trehalose accumulation were proposed. System I would require the UDPG-linked trehalose synthase, whereas system II would constitute an alternative pathway, specifically induced or activated by the expression of a MAL gene. The presence of system II in its constitutive form in the constructed diploids would favour trehalose synthesis during growth on glucose, however, it did not overcome the glycogen deficiency during sporulation nor the lack of trehalose for germination. It seems that only system I, namely trehalose 6-P-synthase, plays a role in the germination process.
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    Protein secretion in yeast: Two chromosomal mutants that oversecrete killer toxin in Saccharomyces cerevisiae (1983)
    Springer
    Current genetics 7 (1983), S. 449-456 
    Details
    ISSN: 1432-0983
    Keywords: Oversecretion mutants ; Protease defect ; Wall glucan defect ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two chromosomal mutations in yeast that result in oversecretion of the K1 killer toxin protein were examined. A recessive mutation in gene ski5 appears to lead to toxin oversecretion through a defect in a cell surface, PMSF-inhibited protease. A wild type killer strain degraded toxin following synthesis, and degradation could be partially prevented by addition of PMSF to the growth medium. The ski5 mutation caused an approximate ten fold oversecretion of toxin, similar to that seen in a PMSF-treated wild type culture, and no increased oversecretion in the presence of PMSF. The ski5 mutation caused oversecretion of other low molecular weight secreted proteins and appeared to oversecrete the α-factor pheromone, as judged by activity tests. The ski5 mutation was complemented by mutations in ski genes 1–4, and the mutant was not supersensitive to mating pheromones or K2 killer toxin. We also examined killer strains with a mutation in the nuclear gene krel which results in a defective (1→6)-β-D-glucan cell wall receptor for killer toxin. Such strains oversecrete toxin into the growth medium, but also, unexpectedly, oversecrete most other secreted proteins. The defect in (1→6)-β-D-glucan in these mutants appears to perturb the partitioning of secreted proteins between the cell wall and the medium.
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    Analysis of yeast DNA by alkaline filter elution (1983)
    Springer
    Current genetics 7 (1983), S. 427-431 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; DNA ; Alkaline elution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The method of analysis of DNA in mammalian cells by alkaline elution from filters (Kohn et al. 1974) was adapted for studies on yeast DNA. By this technique spheroplasts obtained from yeast cells are lysed on filters and single-stranded DNA fragments selectively eluted by alkaline solutions. The procedure was applied to monitor the occurrence of replication intermediates and production of DNA single-strand breakage by MMS, and its repair in growth medium.
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    The study of a rDNA replicator in Saccharomyces (1983)
    Springer
    Current genetics 7 (1983), S. 433-438 
    Details
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Yeast transformation ; Yeast autonomously replicating sequences ; Ribosomal RNA genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have previously demonstrated that the loss of Rcp-CEN3, a centromeric plasmid containing yeast rDNA autonomously replicating sequences (ARS) is as high as around 50% per generation for most yeast strains. In this study we have attempted to elucidate mechanisms underlying the high mitotic instability of Rcp-CEN3. For this purpose a tandem duplication of a rDNA ARS was constructed in Rcp-CEN3. The new plasmid having two ARSs possesses a markedly higher mitotic stability as compared to a monoARS Rcp-CEN3. The mitotic stability of this centromere-containing plasmid which has two replicators corresponds to the calculated value for the mitotic stability of two monoARS plasmids Rcp-CEN3 in given cells. Genetic analysis has demonstrated that both plasmids having one or two ARSs are maintained in the single copy state. These results demonstrate that the mitotic instability of centromeric plasmid Rcp-CEN3 carrying a rDNA ARS is associated with the absence of stringent control of replication from the rDNA ARS. A possible mechanism of replication of the chromosomal rDNA repeats in yeast is discussed in the light of this data.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00377608
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    Electronic Resource
    Electronic Resource
    Mating-type-specific cell cycle arrest and shmoo formation by α pheromone of Saccharomyces cerevisiae in Hansenula wingei (1983)
    Springer
    Archives of microbiology 136 (1983), S. 79-80 
    Details
    ISSN: 1432-072X
    Keywords: α Pheromone ; Cell cycle ; G1 arrest ; Hansenula wingei ; Saccharomyces cerevisiae ; Shmoo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cell cycle of a (5) mating type cells of Hansenula wingei was arrested in the G1 phase by α pheromone of Saccharomyces cerevisiae but not by α(21) pheromone of H. wingei, although both the α pheromones are known to induce sexual agglutination ability of a mating type cells of H. wingei. Cells of α mating type of H. wingei became shmooed or arrested in the G1 phase in response to neither a pheromone of H. wingei nor α pheromone of S. cerevisiae.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00415615
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