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  • 1
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Mutation of bimG, the major protein phosphatase 1 gene in Aspergillus nidulans, causes multiple cell cycle and hyphal growth defects that are associated with overphosphorylation of subcellular components. We have used functional translational fusions with the green fluorescent protein (GFP) to show that BIMG has at least four discrete locations within growing hyphae. Three of these locations, the hyphal tip, the spindle pole body and the nucleus, correlate with previously known requirements for bimGPP1 in mitosis and hyphal growth and are highly dynamic. BIMG-GFP in the hyphal tip seemed to be associated with the plasma membrane and formed a collar of fluorescence within the apical dome. The distribution of nuclear BIMG-GFP varied depending on nutritional conditions; on poor medium, it concentrated more in the nucleolus than in the nucleoplasm, whereas on rich medium, it was more evenly distributed between the two nuclear regions. The association of BIMG-GFP with developing septa was transient, and we present evidence that BIMG phosphatase plays a direct role in septum formation, distinct from its role in mitosis. We conclude that, by being physically present at several sites, the BIMG phosphatase has roles in multiple cellular processes.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Calcium signalling is little understood in filamentous fungi largely because easy and routine methods for calcium measurement in living hyphae have previously been unavailable. We have developed the recombinant aequorin method for this purpose. High levels of aequorin expression were obtained in Neurospora crassa, Aspergillus niger and Aspergillus awamori by codon optimization of the aequorin gene. Three external stimuli (mechanical perturbation, hypo-osmotic shock and high external calcium) were found transiently to increase [Ca2+]c. Each of the calcium signatures associated with these physico-chemical treatments was unique, suggesting the involvement of three distinct calcium-mediated signal transduction pathways. The fungal calcium channel blocker KP4 inhibited the [Ca2+]c responses to hypo-osmotic shock and high external calcium, but not to mechanical perturbation. The divalent cation chelator BAPTA inhibited [Ca2+]c responses to mechanical perturbation and hypo-osmotic shock. The calcium agonists A23187 and cyclopiazonic acid increased [Ca2+]c levels.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 346 (1990), S. 769-771 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Fluo-3 is a fluorescent Ca2+ indicator which has been successfully used to monitor changes in cytoplasmic Ca2+ levels ([Ca2+]j) in animal cells7. Increases in [Ca2+]j lead to increases in Fluo-3 fluorescence intensity. This allows approximate calibration of [Ca2+]i (refs 5, 6). The dye does not ...
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 358 (1992), S. 753-755 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Wheat leaf protoplasts were loaded with Fluo-3 and caged Ca2+ or caged inositol 1,4,5-trisphosphate (InsP3)8"10, and cytosolic free Ca2+ ([Ca2+]i) imaged using laser scanning con-focal microscopy (LSCM)11. The caged probes were photoacti-vated with ultraviolet light. [Ca2+]j was elevated to 〉1 u,M ...
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2048
    Keywords: Cell surfaces ; Cryofixation ; Freezing (extracellular, intracellular) ; Ice deposit (leaf) ; Intercellular space ; Leaf (water, ice) ; Phaseolus (leaf, cryofixation) ; Water droplet (artefacts, leaf)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An experimental study is described of the formation of extracellular deposits on the surfaces of cells in freeze-fractured, frozen-hydrated primary leaves of Phaseolus vulgaris examined by low-temperature scanning electron microscopy. The deposits, observed under a range of experimental conditions, consisted of (a) droplets with diameters of 1.5 to 3.0 μm, (b) droplets with diameters of 10 to 30 μm, (c) crystals with diameters of 1.0 to 6.0 μm, and (d) granules with diameters up to 0.15 μm. The types of deposit were influenced by specimen cooling rate, and their distribution was influenced by the direction of the thermal gradient during cooling. All deposits were predominantly water ice. The quantities of deposited water (up to 4.0% of the leaf water content) increased as the cooling rate was reduced. It is concluded that the ice deposits were primarily artefacts of cryofixation and do not represent the location of water in vivo, as recently suggested. We propose that the deposits arose in four main ways: (1) displacement of water from underlying cells by a pressure wave resulting from the volume increase of intracellular water as it freezes, (2) evaporation of water from warmer cells and its condensation onto colder cells, (3) withdrawal of water from underlying cells by extracellular ice crystallization, (4) condensation of pre-existing water vapour in the intercellular spaces onto cells. The significance of the findings is discussed in relation to the use of lowtemperature scanning electron microscopy in studies of plant morphology and for localizing water and soluble ions within plant cells and tissues.
    Type of Medium: Electronic Resource
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