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  • Electron microscopy
  • Saccharomyces cerevisiae
  • Springer  (40)
  • 2020-2024
  • 1990-1994
  • 1980-1984  (40)
  • 1984  (40)
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Publisher
Years
  • 2020-2024
  • 1990-1994
  • 1980-1984  (40)
Year
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular life sciences 40 (1984), S. 1159-1161 
    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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  • 2
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    Current genetics 8 (1984), S. 559-566 
    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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  • 3
    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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  • 4
    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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  • 5
    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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  • 6
    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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  • 7
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    Current genetics 9 (1984), S. 107-111 
    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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  • 8
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    Current genetics 8 (1984), S. 81-84 
    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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  • 9
    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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  • 10
    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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  • 11
    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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  • 12
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    Current genetics 8 (1984), S. 575-580 
    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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  • 13
    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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  • 14
    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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  • 15
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    Archives of microbiology 137 (1984), S. 357-361 
    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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  • 16
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    Archives of microbiology 138 (1984), S. 273-277 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Secondary metabolite ; Allelopathy ; Photosynthesis ; Electron transport ; Thylakoids ; Herbicides ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cyanobacterin is a secondary metabolite produced by the cyanobacterium, Scytonema hofmanni. Highly purified cyanobacterin was found to inhibit the growth of many cyanobacteria at a minimum effective dose of 2 μg/ml (4.6 μM). The antibiotic had no effect on eubacteria including the photosynthetic Rhodospirillum rubrum. The site of action of cyanobacterin was further investigated in the unicellular cyanobacterium, Synechococcus sp. Electron micrographs of antibiotic-treated Synechococcus cells indicated that cyanobacterin affects thylakoid membrane structure. The antibiotic also inhibited light-dependent oxygen evolution in Synechococcus cells and in spheroplasts. These data support our conclusion that cyanobacterin specifically inhibits photosynthetic electron transport. This activity is similar to herbicides such as 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU). The anhydro analog of cyanobacterin had no biological activity.
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  • 17
    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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  • 18
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    Archives of microbiology 140 (1984), S. 265-270 
    ISSN: 1432-072X
    Keywords: EcoRI ; EcoRI-DNA complexes ; EcoRI* activity ; Recognition sites ; Frequency of binding ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Electron microscopy of negatively stained isolated restriction enzyme EcoRI revealed particle projections with triangular or square outlines, indicating that the enzyme, in its tetrameric state, is tetrahedron-like. The two dimers making up the tetramer appear to be arranged in two planes orthogonal to each other. Complexes formed by EcoRI with the plasmids pBR322 or pGW10 were investigated by electron microscopic spreading techniques. In the presence of Mg2+, EcoRI was bound to the DNA molecules to form pearl necklace-like aggregates. The number of bound EcoRI particles was much higher as the sum of EcoRI-and 5′..AATT..3′ sites (with exceptions, the 5′..AATT..3′ sites may function as one type of EcoRI* sites) along the DNAs, indicating unspecific binding. In the absence of Mg2+, EcoRI was bound to the DNA only at the recognition site for EcoRI and the sites where the tetranucleotide sequence 5′..AATT..3′ was present. A direct correlation of the local concentrations of the bases A and T within the flanking sequences of the binding sites with the frequency of EcoRI to the DNA was observed. Dimers and tetramers of the enzyme was found to bind to the DNA. Tetramers occasionally exhibited two binding sites for DNA as indicated by the observation of DNA loops originating at the sites of bound tetrameric EcoRI particles.
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  • 19
    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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  • 20
    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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  • 21
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    Cell & tissue research 237 (1984), S. 219-226 
    ISSN: 1432-0878
    Keywords: Spermatogonia ; Spermatocytes ; Carbohydrates ; Guppy ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The structure of guppy (Poecilia reticulata) spermatogonia and spermatocytes has been studied using electron microscopy. The spermatogonia, situated at the apex of the seminiferous tubule, are almost all surrounded by a network of Sertoli cells; they have very diffuse chromatin and one or two large nucleoli. The cytoplasm contains relatively few organelles, although annulate lamellae are found. The mitochondria have few cristae and are concentrated at one pole of the cell; they are sometimes found with intermitochondrial cement. These spermatogonia are separated from each other, having no intercellular bridges or inclusion in Sertoli cells, and are relatively undifferentiated; they correspond to stem cells. The spermatogonia beneath the apex are organized into cysts. First-generation spermatogonia are more dense and heterogeneous, their nuclei becoming smaller and their chromatin becoming denser during successive generations. In spermatocytes, the synaptinemal complex exists as a modified form until metaphase. The concentration of organelles in the cytoplasm increases and the organelles become more diversified as spermatogenesis progresses. Many cytoplasmic bridges are observed (several per cell), indicating that the cells remain in contact after several divisions. These changes in germ cell structure have been related to some of the characteristic features of spermatogenesis in guppy, e.g. the large number of spermatogonial generations and the complexity of spermiogenesis.
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  • 22
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    Cell & tissue research 238 (1984), S. 413-416 
    ISSN: 1432-0878
    Keywords: Peroxisomes ; DAB-cytochemistry ; Electron microscopy ; Liver, amphibian ; Gymnophiona ; Ichthyophis glutinosus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of hepatic peroxisomes was investigated in Ichthyophis glutinosus (Amphibia: Gymnophiona), employing perfusion fixation and the diaminobenzidine (DAB) technique for the visualization of catalase. The majority of peroxisomes is circular or rod-shaped, although elongated particles occasionally occur. They contain a finely granular matrix, lightly stained after the DAB procedure. Their mean diameter is approximately 0.25 μm. Serial sections reveal that the circular and rod-shaped peroxisomal profiles are cross and oblique sections of highly tortuous, tubular organelles exceeding 2 μm in length. In addition to tubular profiles, elongated, rectangular particles, as well as straight dumbbell-shaped organelles with distinct marginal plates are observed. They range from 900 to 1650 nm in length (mean = 1200 nm). In the flattened, thin central portion of the dumbbell-shaped particle, the peroxisomal membranes form a cisterna enclosing one or two uniformly thick marginal plates, which display a definite substructure with a periodicity of 10 nm. These findings indicate that peroxisomes in the liver of Ichthyophis exhibit a complex organization. It is suggested that the organelles undergo a specific differentiation process, morphologically characterized by the formation of enlarged segments of unusual shape.
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  • 23
    ISSN: 1432-0878
    Keywords: Ovulation (rabbit) ; Graafian follicle ; Perfusion ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Isolated ovaries from untreated, sexually mature rabbits were introduced into an in vitro perfusion system and perfused with a chemically defined medium containing albumin. The ovaries were perfused for up to 15 h (mean 11.5 h) and then processed for morphological investigation. Both at the light- and electron-microscopical levels, most of the ovaries exhibited a normal structure comparable with ovaries in situ. In two cases, however, marked accumulations of bacteria were found, although not inside the follicles. Since ovulation in the rabbit normally occurs between 9.5–13 h after mating or human chorionic gonadotrophin treatment, this model seems adequate for studies of ovulation in vitro. It is, however, important to study the ovaries microscopically after the perfusion to detect artifacts, e.g., bacterial infection, that may have influence on the process of ovulation.
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  • 24
    ISSN: 1432-0878
    Keywords: Tannic acid ; Acetylcholine receptors ; Tissue culture ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Spinal cord neurons from 9-day chick embryos were maintained in culture for up to 35 days and then fixed in 4% cacodylate-buffered glutaraldehyde containing 2% tannic acid. After about 15 days in culture a small percentage of the synaptic specializations present were characterized by striking electron-dense striations averaging 15 nm in width, oriented perpendicular to the postsynaptic membrane. These structures increased in frequency with time in culture (to a maximum of about 10% of all synapses in the oldest cultures); they were asymmetrical, protruding approximately 8 nm into the synaptic cleft, and more deeply (approximately 15–18 nm), into the postsynaptic cytoplasm. On the basis of earlier work by Sealock (1980) they are interpreted as concentrations of acetylcholine receptors. Similar membrane differentiations were also seen associated with active-zone areas of a few presynaptic membranes, and the possibility that these represent presynaptic acetylcholine receptors is discussed. Additional observations reported are (1) the presence of striations resembling those seen at the postsynaptic membrane in the membranes of some postsynaptic vesicles, and (2) filamentous links between the striations and cytoskeletal elements of the postsynaptic cell.
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  • 25
    ISSN: 1432-0878
    Keywords: Monoamine-containing cells ; Taste bud ; Paracrine cells ; Mechanoreceptors ; Electron microscopy ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The taste buds on the barbels in three species of teleosts (Cyprinus carpio, Misgurnus anguillicaudatus, Parasilurus asotus) were studied by means of fluorescence and electron microscopy. Intensely yellow-fluorescent cells, which are disk-shaped and located exclusively in a basal position, are observed in the barbel-buds of all fishes examined. The basal cells contain a large number of small clear vesicles approximately 40–60 nm in diameter, which show a tendency to aggregate in the cytoplasm facing the junction of the nerve terminals; chemically transmitting synapses are seen in the latter region. It is suggested from the present observations that the basal cells in the barbel-bud may originate from Schwann cells and have a dual function both as mechanoreceptors and paracrine elements. Since the administration of 5,6-DHT results in an appearance of small dense vesicles among the small clear vesicles, the possibility exists that the basal cell may be capable of taking up monoamines and storing them in the small clear vesicles.
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  • 26
    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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  • 27
    ISSN: 1432-0878
    Keywords: Chloride cells ; Acid stress ; Gill ; Electron microscopy ; Fathead minnow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Fathead minnows, Pimephales promelas, were exposed for 129 days to Lake Superior water acidified with sulfuric acid by means of a flow-through toxicant injection system. The effects of chronic acid stress (pH 6.5, 6.0, 5.5, 5.0) on gill histology were examined. Most of the histological effects were seen at pH 5.5 and 5.0 and were confined primarily to changes in numbers, distribution, and morphology of chloride cells. At low pH levels there tend to be more chloride cells in the gill epithelium and an increased percentage of these cells in the secondary lamellae. In contrast to normal chloride cells, chloride cells from fish exposed to low pH frequently had apical pits while some had bulbous apical evaginations. The occurrence of structural changes in chloride cells during exposure to acid water suggests that chloride cells may be involved in acclimation to acid stress.
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  • 28
    ISSN: 1432-0878
    Keywords: Lactating cell ; Lipid droplets ; Secretory vesicles ; Mitochondria ; Intracellular associations ; Electron microscopy ; Milk secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The size, cellular location, and identity of surface-associated components were determined for lipid droplets in lactating cells. Transmission electron-microscopic measurements were made involving 3801 droplets in approximately 211 cells from three rats and 1197 droplets in 66 cells from a mouse. For the purposes of droplet evaluation, cells were divided into seven locations ranging from basal to secreting positions. Droplets were also categorized with respect to contact with other droplets, basolateral plasma membrane, mitochondria, Golgi apparatus, secretory vesicles, and endoplasmic reticulum-cytoplasm (ERC). Data on droplet size showed that droplet growth occurs mainly in the secretory position, confirming previously published findings. Lipid droplets from mouse tissue, although somewhat smaller in size showed similar growth trends to those of the rat. Data on numbers of droplet contacts and percentages of droplet circumferences involved in associations with other cell components showed that the dominant interaction of lipid droplets was with the ERC. However, intimate association of droplets with mitochondria was noted in all cellular locations. In addition, nursed animals exhibited a greater proportion of droplet surface association with secretory vesicles and less in contact with mitochondria in comparison to those not nursed. The significance of these relationships to milk synthesis and secretion is discussed.
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  • 29
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    Cell & tissue research 236 (1984), S. 249-255 
    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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  • 30
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    Cell & tissue research 235 (1984), S. 51-58 
    ISSN: 1432-0878
    Keywords: Parathyroid glands ; Electron microscopy ; Light microscopy ; Quantitative histology ; Mongolian gerbil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Stereology and semi-automatic image analysis were used with the aim of comparing the structure of parathyroid glands from untreated adult Mongolian gerbils fixed by immersion with those fixed by perfusion. Subclassification of the chief cells based upon the staining affinity or electron density of the cytoplasm was readily performed only in glands fixed by immersion, and so-called atrophic cells were observed only in these glands. The atrophic cells were often surrounded by “light” chief cells. In glands fixed by perfusion, “light” chief cells were only rarely encountered. A significant difference between glands fixed by immersion and those fixed by perfusion was found only with regard to the form of cells and nuclei, those fixed by perfusion being more spherical. When comparing individual cells within glands fixed by immersion, “light” chief cells were more spherical and had a significantly larger nuclear and cellular size, and a lower mitochondrial volume density than the “intermediate”/“dark” chief cells. Otherwise there were no significant differences in any of the parameters investigated. These data indicate that occurrence of socalled “light” chief cells and atrophic cells is a result of improper fixation. The results of this study do not favour the concept of a functional cycle with a simultaneous occurrence of active and inactive cells within parathyroid glands.
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  • 31
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    Cell & tissue research 235 (1984), S. 153-158 
    ISSN: 1432-0878
    Keywords: Bovine ; Interdigitating cell ; Lymphoid tissues ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Electron-microscopic studies of lymphoid tissues from bovine fetuses and from calves disclosed a non-lymphoid cell type in the thymus-dependent zones of secondary lymphoid tissues and in the thymus that is distinguishable from reticulum cells, epithelial and endothelial cells, and macrophages. Based on morphological and topographical criteria, the cell is identified as the interdigitating cell. In addition, studies of the tissues of normal and virus-challenged fetuses, and of conventionally reared calves, indicated that the interdigitating cells originate from monocytoid cells, which undergo differentiation in the thymus-dependent zones during an immune response.
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  • 32
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    Cell & tissue research 235 (1984), S. 243-249 
    ISSN: 1432-0878
    Keywords: Gap junction ; Electron microscopy ; Freeze fracture ; Cell-to-cell communication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Gap junctions exist in the septa between the segments of the lateral giant axons in the ventral nerve cord of the crayfish Procambarus. A large increase in the resistance (uncoupling) of these gap junctions was brought about by mechanical injury to the axonal segments. Both thin sections and freeze-fracture preparations were used to monitor the morphological changes which occurred up to 45 min after injury. There was no apparent change in the organization (a loose polygonal array) of the intramembrane particles which make up the junctional complex up to 45 min after injury. In some instances, however, the intramembrane particles appeared to have moved away from the junctional area. Other junctional regions were internalized and appeared similar to what have been called annular gap junctions. Also at this time (20–25 min after injury), a dense cytoplasmic plug formed in uninjured axon near the junctional region. It is concluded that the gap junctions that exhibit a loose polygonal organization of the intramembrane particles may be either in a state of low resistance (coupled) or a state of high resistance (uncoupled).
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  • 33
    ISSN: 1432-0878
    Keywords: Ovulation ; Perfusion ; Graafian follicle (rabbit) ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ovulation was induced in rabbits by intravenous administration of human chorionic gonadotrophin (HCG), and 4–5 h later the ovaries were isolated and introduced into an in-vitro perfusion system containing synthetic medium with albumin. Rupture of follicles occurred in vitro within the physiological time range (mean 11.3 h after injection of HCG), although with a reduced frequency. Preovulatory and ruptured follicles were studied in detail by light and electron microscopy. In the granulosa layer of ruptured or preovulatory follicles cytoplasmic blebbing activity, disappearance of CallExner bodies and differentiation toward luteinized cells were found. Perhaps the most important sign of normal preovulatory development in vitro was that the basement membrane surrounding the granulosa layer was penetrated by projections of granulosa cells. In the absence of this penetration phenomenon the granulosa layer prolapsed out of the follicle. Immediately before rupture, follicles showed marked degeneration, restricted to the outer layers of the apical wall, which is compatible with the hypothesis that degradative enzymes are released close to the surface of preovulatory follicles. Although the majority of follicles that ovulated under in-vitro conditions showed the same kind of morphological alterations as can be seen in vivo, occasional atypical ruptures occurred without any overt signs during perfusion. Also technical manipulations of the perfusion system, e.g., nonphysiological increase of perfusion pressure, could force follicles to rupture. This illustrates the importance of careful morphological study of all ovaries perfused in vitro before conclusions are drawn.
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  • 34
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    Cell & tissue research 237 (1984), S. 149-154 
    ISSN: 1432-0878
    Keywords: Pituitary gland, pars distalis ; Innervation ; Synaptoid contacts ; Electron microscopy ; Rana temporaria L
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the pars distalis of the hypophysis of adult Rana temporaria, three types of nerve-fiber profiles were found at two distinct sites, in both lateral parts of the bordering regions of the anterior lobe with the intermediate lobe of the hypophysis. The first type of nerve-fiber profile consists of bundles of very fine axonal elements (diameter: 〈0.7 μm). The second type is formed by larger nerve fibers (diameter up to 4 μm) containing a few neurosecretory granules of approximately 100 nm. The third type of nervefiber profile resembles the second type but these nerve fibers make synaptoid contacts on at least two different types of glandular cells. The possible functional significance of these nerve fibers in the pars distalis is discussed. No nerve fibers were found (1) in the central part of the bordering region of the pars distalis with the intermediate lobe, (2) at the bordering region with the median eminence and (3) with the neurohypophysial stalk, and (4) in all other parts of the pars distalis.
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  • 35
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    Cell & tissue research 238 (1984), S. 165-175 
    ISSN: 1432-0878
    Keywords: Mesonephros ; Pig embryo ; Glomerulus ; Microvasculature ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Vascularization of the pig mesonephros was investigated in embryos 5–8 cm in length. Vascular injections with microfil were cleared and dissected; corrosion casts were studied under the scanning electron microscope (SEM). Perfusion-fixed tissue was used for SEM and transmission electron microscope (TEM) studies, including freeze-fracture specimens. The branches of one mesonephric artery carry up to 15 glomeruli. Several glomeruli occupy the same arterial branch, with very short afferent arterioles proper. The efferent vessels, frequently 2–5, leave the extensive vascular pole opposite the entering arteriole and split into peritubular capillaries radiating towards the superficial veins. These capillaries form vascular regions in the shape of flattened pyramids. Along its course, one nephron is supplied by vessels derived from 4–7 glomeruli. The nephrons have less vascular contact than in the definitive kidney. The ultrastructure of the single mesonephric vessels matches the metanephric counterparts. Epithelioid cells with renin granules are common in afferent arterioles, larger arteries, and efferent vessels. The lobulated glomeruli are up to 750 μm long and flattened, showing the usual features of podocytes, mesangial cells, and an attenuated endothelium with fenestrations between 50 and 250 μm. It partially retains its own basement membrane. There is no proximal mesangium.
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  • 36
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    Cell & tissue research 238 (1984), S. 387-394 
    ISSN: 1432-0878
    Keywords: Pacinian corpuscles, cat ; Denervation ; Atrophy ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of Pacinian corpuscles of the cat located in the crural region and innervated by the interosseous nerve was studied 1 to 14 months after denervation. Both the Pacinian inner core and capsule remained well preserved one month after denervation. However, the denervated inner cores underwent progressive atrophy and wasting, which resulted in a gradual reduction of the amount of inner-core cells and lamellae, widening of interlamellar clefts, formation of empty spaces in the axial region and a considerable increase in the number of collagen fibrils. In spite of the wasting, the inner core still survived 14 months after denervation, but at least half of its volume became occupied by collagen fibrils which surrounded the remaining inner-core cells and lamellae. Collagen fibrils assembled in the denervated core were markedly thinner than those found in the capsule, as is also the case in normal Pacinian corpuscles. In the capsule, discrete focal degeneration, occasional pyknosis of the innermost capsular cells and macrophage infiltration were observed from the first month after nerve section onward, but the number of capsular layers remained within the normal range (30–40) up to 14 months after denervation.
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  • 37
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    Protoplasma 119 (1984), S. 48-54 
    ISSN: 1615-6102
    Keywords: Electron microscopy ; Marine alga ; Membrane contrast ; Osmium-azole complexes ; X-ray microanalysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Addition of certain heterocyclic nitrogen-carbon compounds to standard osmium tetroxide solutions used as secondary fixative resulted in an enhanced general membrane contrast in cells of the marine algaEmiliania huxleyi (Lohmann) Kamptner. Ultrastructural cell morphology and the contrast distribution were compared between cells treated according to a standard secondary fixation procedure and cells post-fixed when above mentioned heterocyclic compounds were introduced; in both cases some of the ultrathin sections were post-stained. Different compounds were tested: 1,2,4-triazole (TRA), 3-amino-1,2,4-triazole (A-TRA), 5-amino-tetrazole (A-TEA) and 2,4,6-tri-amino-1,3,5-triazine (melamine). The results were interpreted to indicate the possible bonding types arising from interaction of the heterocyclic compounds with osmium tetroxide and with membrane constituents. Interpretations were partly inspired by considerations from coordination chemistry. All above tests which did not include post-staining of thin sections could be performed at alkaline pH, and consequently calcified structures were preserved. The enhanced osmium accumulation at membranes was verified with X-ray microanalysis, which also showed that in the cases where membranes were visibly contrasted, localization of probable sites of intracellular non-crystalline calcium was facilitated.
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  • 38
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    Plant and soil 76 (1984), S. 127-137 
    ISSN: 1573-5036
    Keywords: Adenylate pool ; Biomass volume ; CO2 evolution ; Chitin ; DNA ; Electron microscopy ; Enzymes ; Fluorescent antibody ; Fumigation-respiration ; Fungi Histochemistry ; Imunofluorecence ; Jones-Mollison technique ; Microcosms ; Monoclonal antibodies ; Nitrogen ; Nutrients ; Oxygen consumption ; Phosphorus ; Phytotoxins ; Plate counts ; Rhizobium ; Rhizosphere ; Sulphur ; Xenobiotics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary There is an immense literature on biological and biochemical analyses of soils. Such analyses have revealed the enormous richness of species in soil and their vast range of metabolic potentials and ecological diversity. Accordingly, the approaches used to investigate the soil biota and its biochemistry usually have to be modified or adapted depending upon the purpose of the investigation. Studies of micro-organisms in the soil environment, are complicated because microbial cells are commonly attached to surfaces where they live side-by-side with other populations in consortia usually containing different morphological and physiological types. Such assemblages of organisms cannot be described quantitatively using cultural techniques, such as plate counts, which underestimate both cell numbers and viable biomass. The development of more powerful observational and staining techniques has improved our knowledge of the diverse morphological and biochemical composition of soil micro-communities. Such findings have been amplified at a grosser level by laboratory studies with multi-component systems (microcosms) to mimic field situations and to assess the range of biochemical potentials of microbial consortia. But despite notable advances in analytical methods we are still, with a few exceptions, unable to detect or identify those microorganisms which carry out specific biochemical transformations or determine whether particular cells are alive, dormant or dead at the time of observation. Considerable work has been done to define some of the fundamental ecological attributes of microbial assemblages in soil. Productive work on the metabolic activities of the soil microbiota, specially geochemical transformations of C, N, S and P, has been under way for more than a century. But only in more recent years have more sensitive and reproducible analytical methods become available to measure viable biomass in soil. This will enable some insight to be gained into the role that microbial biomass plays as a labile source and sink for plant nutrients.
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  • 39
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    Plant and soil 76 (1984), S. 319-337 
    ISSN: 1573-5036
    Keywords: Aggregates ; Aluminium ; Bacterial mucilage ; Binding agents ; Calcium ; Cation bridges ; Complexing agents ; Dispersion ; Electron microscopy ; Electrophoretic mobility ; Fungal hyphae ; Glues Iron ; Management Periodate ; Polysaccharides ; Rhizosphere ; Roots ; Slaking
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The stability of pores and particles is essential for optimum growth of plants. Two categories of aggregates macro- (〉 250 μm) and micro- (〈250 μm) depend on organic matter for stability against disruptive forces caused by rapid wetting. Dispersion of clay particles from microaggregates is promoted by adsorption of complexing organic acids which increase the negative charge on clays. The acids are produced by plants, bacteria and fungi. However, the dispersibility of clay in microaggregates is offset by the binding action of polysaccharides, mainly mucilages produced by bacteria, but also by plant roots and fungal hyphae. The stability of microaggregates is also enhanced by multivalent cations which act as bridges between organic colloids and clays. Macroaggregates are enmeshed by plant roots, both living and decomposing, and are thus sensitive to management, and increase in number when grasses are grown and the soil is not disturbed. Lack of root growth,i.e. fallow, has the opposite effect. Various implications for management of soil structure are discussed.
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  • 40
    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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