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  • 1
    ISSN: 1615-6102
    Keywords: Actin-binding protein ; Green algae ; Immunolocalization ; Micrasterias denticulata ; Profilin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Profilin is detected by means of immunoblotting in the green algaMicrasterias denticulata at a molecular mass of about 14 kDa with antibodies against celery root profilin, recombinant tobacco profilin, and recombinant birch profilin. Poly-L-proline purification ofM. denticulata extracts leads to a single band at 14 kDa. By means of immunoelectron microscopy first evidence is provided for the presence of profilin in a microtubule center associated with the migrating nucleus in high-pressure-frozen and freeze-substituted cells. Colocalization with a particular filamen-tous-actin aggregation in the same area suggests a role of profilin in the process of nuclear migration. Moreover staining is visible in several areas of the nucleus including the nucleolus, heterochromatin, and nuclear-pore complexes. An even distribution of profilin is found throughout the cytoplasm at the confocal-microscopy level as well as by immunogold labeling. Occasionally also areas at the surface of the chloroplast are stained.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1615-6102
    Keywords: Actin ; Actin-binding proteins ; Gelsolin ; Micrasterias ; Microinjection ; Profilin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Recombinant profilins from different sources (Betula verrucosa, Schizosaccharomyces pombe, Acanthamoeba castellani, or man) cause marked effects on cell growth and morphogenesis when microinjected into growing cells of the green algaMicrasterias denticulata. Whereas control injections with β-lactoglobulin only result in a slight delay of cell growth, when profilin is injected cell differentiation ceases and only resumes about 1 to 2 h after the injection, depending on the dose. The resulting cell does not show any malformations, but is reduced in size and retarded in differentiation compared to controls. As a consequence of the profilin microinjection the pattern of cytoplasmic streaming and cytoplasmic structure are also altered. Gelsolin, injected for comparison, leads to minor retardation of cell development but produces less marked effects than profilin. Microinjection of fluorescently labeled profilin shows even distribution throughout the cytoplasm and more intense fluorescence in the nucleus. Electron microscopical investigations of cells fixed immediately after profilin injection show a normal distribution of dictyosomes, ER cisternae, microtubules, and secretory vesicles compared to noninjected controls at the same developmental stage. Our results indicate that disturbance of the natural actin turnover by the injection of actin-binding proteins strongly affects development ofMicrasterias, corroborating a key role of actin in the morphogenetic process.
    Type of Medium: Electronic Resource
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