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  • Morphogenesis  (2)
  • Ultraviolet microbeam  (2)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 149 (1989), S. 130-143 
    ISSN: 1615-6102
    Keywords: Colchicine ; Cytochalasin D ; Diatom ; Microtubule center ; Morphogenesis ; Valve morphology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The effects of various drugs on cell wall (valve) morphogenesis was investigated in three species of diatoms (Pinnularia spp., Surirella robusta, andHantzschia amphioxys) using light microscopy (LM) and scanning electron microscopy (SEM). Treatment ofSurirella with the microtubule (MT) disrupting agent colchicine during early valve formation results in a characteristic malformation of the valve, whereby part of the normally circumferential raphe canal forms as an abnormal protruding lip on the valve surface, located up to ∼ 20 μm from the edge of the valve. The position of this malformed lip coincides with the location of a microtubule center (MC) at the time of colchicine addition, suggesting that the MC may play a direct role in positioning the tip of the raphe canal during valve formation. The migration of this MC to the tip of the cell during early valve morphogenesis is reversibly inhibited by the metabolic inhibitor 2-4-dinitrophenol (DNP). The effect of colchicine onPinnularia valve formation is less severe, causing occasional malformation of the raphe, but little if any lateral displacement. InHantzschia, colchicine has no effect on the positioning of the raphe, but prolonged exposure causes fusion of the raphe canal with the valve face. Cochicine treatment also results in the absence of the normal curvature at the central interruption in the raphe, as well as abnormal pore formation in this central area. Addition of cytochalasin D during early valve formation inHantzschia causes the raphe canal to form in the center of the valve face, suggesting that the normal translocation of the raphe canal to the valve edge is actindependent. Comparison of valves from control and cytochalasintreatmentHantzschia suggest that the pore spacing within the valve is determined by the position relative to the raphe, and does not depend on whether to pores form on the side (mantle) or the face of the mature valve.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1615-6102
    Keywords: Mitosis ; Ultraviolet microbeam
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We describe the assembly of a UV microbeam microscope based on a Zeiss IM35 inverted microscope. The important UV transmitting elements are standard UV epifluorescence attachments available from Zeiss; the main modification involves fitting an adjustable slit in place of the field diaphragm. We describe how to align and focus the UV source for optimal irradiations. Our current version of this machine is also fitted with a monochromator and using monochromatic UV light, we can reproduceably create Areas of Reduced Birefringence in spindle fibres with ca. 2–3 s irradiations, while continually observing the fibres. The microscope is stable and easy to set up, allowing many consecutive experiments to be done, including multiple irradiations on the one cell. In conjunction with video image processing techniques, the cells can be observed continuously using polarising, Nomarski or other optical systems. Some preliminary observations demonstrating the versatility of the machine are described.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1615-6102
    Keywords: Mitosis ; Ultraviolet microbeam ; Spindle fibres ; Microtubules ; Crane-fly spermatocytes ; Newt epithelial cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In order to resolve apparent differences in reported experiments, we directly compared the effects of ultraviolet (UV) microbeam irradiations on the behaviour of spindle fibres in newt epithelial cells and crane-fly spermatocytes, using the same apparatus for both cell types. This work represents the first time that irradiated crane-fly spermatocytes have been followed using a high-NA objective and video-enhancement of images. In both cell types, irradiation of a kinetochore fibre in metaphase produced an area of reduced birefringence (ARB), known to be devoid of spindle microtubules (MTs). Subsequently the kinetochore-ward edge of the ARB moved poleward with average velocities of 0.5 μm/min (n=20) in spermatocytes and 1.1 μm/min (n=6) in epithelial cells. The poleward edge of the ARB rapidly disappeared when viewed using a ×100, high-NA objective but generally remained visible when viewed with a ×32, low-NA objective; this difference suggests that MTs poleward from the ARB disperse vertically out of the narrow depth of field of the ×100 objective but that many remain encompassed by that of the ×32 objective. The primary difference in response between the two cell types was in the behaviour of the spindle poles after an ARB formed. In spermatocytes the spindle maintained its original length whereas in epithelial cells the pole on the irradiated side very soon moved towards the chromosomes, after which the other pole did the same and a much shortened functional metaphase spindle was formed.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1615-6102
    Keywords: Diatom ; Morphogenesis ; Seta ; Silica ; Spine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary InChaetoceros peruvianus, the two very long, delicately tapered setae (spine-like processes) of the “lower” valve curve downwards gently until they are often almost parallel, while those emerging from the “upper” valve curve sharply downwards until oriented almost in the same direction as the setae of the lower valve. This curvature creates a deep pit between the bases of the upper valve's setae, where they emerge from the valve. In live cells, extension of setae is rapid and very sensitive to disturbance. After cleavage the new silica deposition vesicle (SDV) appears in the centre of the furrow and expands outwards over it. A distinct microtubule centre (MC) appears directly on top of the SDV. Microtubules (MTs) from the MC ensheath the nucleus, and others fan out over the SDV and plasmalemma. A little later, the MC in the lower daughter cell moves off the SDV, and its MTs now appear to mould the plasmalemma/ SDV into the deep pit between the base of the setae. In the upper daughter cell, the MC remains on the SDV. Initiation of setae is first observed as protuberances of bare cytoplasm growing from the sides of the daughter cells, through gaps in the parental valve. Many MTs initially line the plasmalemma of these protuberances as they grow outwards and the SDV also expands over the new surface. As the setae get longer, a unique complex of three organelles appears. Just behind the naked cytoplasm at the tip of the seta, a thin flat layer of fibrous material lines the plasmalemma. This, the first of the complex, is called the “thin band”. Immediately behind this is the second, a much thicker, denser fibrous band, the “thick band”. At the front edge of the SDV, 5–6 “finger-like outgrowths” of silicified wall grow forwards. These are interconnected by the elements of the thick band which thus apparently dictate the polygonal profile of the seta. These also appear to generate the spinules (tiny spines) that adorn the surface of the seta; the spiral pattern of the spinules indicates that this whole complex might differentiate one after the next, in order. Further back from the tip, evenly spaced transverse ribs are formed. These are connected to the third organelle in the complex, the “striated band”; our interpretation is that the striated band sets up the spacing of the ridges that regularly line the inner surface of the setae. During seta growth, this complex is apparently responsible for controlling the delicate tapering curvature of the very fine silica processes. Since the complex is always seen near the tip of the seta, we conclude that it migrates forwards steadily as the tip grows. While the thin and thick bands could slide continuously over the cell membrane, the striated band must be disassembled and then recycled forward during extension if it is indeed connected to the ridges lining the inside of the setae. We could find no indication that turgor pressure drives extension of the setae, in which event the activity of these organelles is responsible for growth using the justformed silica tube as the base from which extension is generated.
    Type of Medium: Electronic Resource
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