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  • 1
    ISSN: 1615-6102
    Keywords: Calcium ; Contraction ; Discophrya ; Ionophore A 23187 ; Ruthenium red ; Tentacle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The suctorian protozoonDiscophrya collini has contractile tentacles with microfilaments and a central microtubule-lined canal (axoneme). The role of calcium fluxing in tentacle contraction has been investigated using the Ca2+ ionophore A 23187 and ruthenium red (RR), a known inhibitor of certain Ca2+ membrane transport events. Treatment with CaCl2 alone caused tentacle contraction with a threshold at 5 × 10−3 M CaCl2 and a maximum at 5 × 10−2 M CaCl2 with contraction to 32.8% of the original length. In the presence of 5 μM ionophore A 23187 the threshold was lowered to 5 × 10−6 M CaCl2 with a maximum to 19.6% original length at 5 × 10−2 M CaCl2. A 23187 alone induced contraction with a threshold of 3.0 μM and a maximum to 30.5% original length at 10 μM A 23187. Treatment with RR alone had little effect on tentacle length. However, a 10 μM A 23187-induced contraction was partially counteracted by the simultaneous application of RR with a threshold at 2 μM RR and a maximum at 8 μM RR. Removal of the ionophore after induced tentacle contraction resulted in partial re-extension, which was inhibited by RR. Ultrastructural observations indicated that the ionophore and CaCl2-induced contraction processes are indistinguishable. The CaCl2/ionophore treatments led to axonome disruption, interpreted as a consequence of supranormal levels of intracellular Ca2+. X-ray microanalysis of cytoplasmic membrane-bound elongate dense bodies (EDB) showed a high level of Ca2+ in CaCl2-treated cells, little Ca2+ in ionophore-treated cells and intermediate levels in the RR-treated, ionophore/RR-treated and untreated control cells. It is suggested that A 23187 enhances both the uptake of extracellular Ca2+ and the release of Ca2+ from the EDB, the latter being counteracted by RR. These observations support the proposal that the EDB act as Ca2+ reservoirs, and that their Ca2+ fluxing moderates cytosolic Ca2+ levels which mediate a Ca2+-dependent tentacle contraction mechanism.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 111 (1982), S. 195-205 
    ISSN: 1615-6102
    Keywords: Discophrya ; Evaginative budding ; Microtubules ; Reproduction ; Suctoria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Discophrya collini reproduces asexually through the formation of a ciliated swarmer by evaginative budding. This process is initiated by the repeated replication of a single subcortical kinetosome to form a kinetosome field. The epiplasm of the multilayered cortex covering this field becomes reduced in thickness and the whole cortex invaginates to produce an internal embryonic cavity. The kinetosomes become organised into rows, and each produces a cilium which projects into the cavity. On completion of the embryonic cavity its walls are extruded through the cavity opening to form an external ciliated swarmer connected to the parent by a thin bridge of cytoplasm. It is suggested that this evagination is induced by a rapid breakdown of supporting microtubules in the cavity wall and the subsequent hydrostatic pressure exerted by the body cytoplasm. The connecting bridge shows no specialised ultrastructural features and separation of swarmer from parent probably is achieved by the active movement of the swarmer. The cytoplasm of the swarmer is similar in structure to that of the adult cell but contains a number of primordia of tentacle axonemes. The infraciliature resembles that of other suctorian swarmers. On settling, the cilia of the swarmer are lost, at least some by resorption, a stalk may be secreted and the axoneme primordia are extended to form functional tentacles.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1615-6102
    Keywords: Discophrya ; Tentacle contraction ; Cations ; Calcium ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Discophrya collini is a suctorian protozoan with contractile tentacles containing a microtubule-lined canal and microfilaments. The effects of a range of cations on tentacle contraction and ultrastructure have been determined. Treatment with 80 mM CaCl2 and 95 mM MgCl2 causes contraction to 28% and 57% of the control length respectively. Re-extension takes over 4 hours in the culture medium, but CaCl2-treated tentacles are re-extended after a 5 minutes treatment with 10−2 M EDTA or 5 × 10−3 M EGTA. CuCl2 causes a significant contraction at 10−5 M (to 77%); LaCl3 at 10−4 M (to 65%); ZnCl2 at 10−2 M (to 65%), but BaCl2, CoCl2, MnCl2, NiCl2, and SrCl2 cause significant changes only at 10−1 M. The cytoplasm of CaCl2-treated cells contains two forms of membraneous structures when viewed in TEM; that of MgCl2-treated cells reveals granular areas of medium electron density. None of these features are seen in control cells. The microtubules of the tentacle canal appear to be intact upon its retraction into the cell with no change occurring in the numbers or relative positions of the microtubules. The tentacle cortex is wrinkled. It is suggested from this and previous work that tentacle contraction may be mediated by a microfilament-based mechanism, and that calcium may be involved.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1615-6102
    Keywords: Calcium ; Contraction ; Discophrya ; Ionophore A23187 ; Ruthenium red ; Tentacles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The tentacles of the suctorian protozoonDiscophrya collini are stimulated to contract by externally applied Ca2+. The role of extracellular Ca2+ in tentacle contraction was studied by monitoring45Ca2+ uptake, using ionophore A23187 to facilitate membrane transport of calcium and ruthenium red (RR) as an inhibitor of transport. The degree of tentacle retraction was dependent upon external Ca2+ concentration and studies with45Ca2+ using scintillation counting indicated a linear relationship between external Ca2+ concentration and Ca2+ uptake. Uptake of Ca2+ was enhanced in the presence of the ionophore while RR caused little inhibition.45Ca2+ uptake was only partially inhibited by RR when cells were subjected to a Ca2+, ionophore and RR mixture. Grain counts from light microscope autoradiographs after treatment of cells with45Ca2+/ionophore,45Ca2+/RR or45Ca2+ alone showed heavy, light and intermediate labelling respectively. In all instances the grains were evenly distributed within the cell. These observations are interpreted as supporting the suggestion that the ionophore enhances both the uptake of extracellular Ca2+ and release of Ca2+from an internal source, while the RR could only partially prevent movement of Ca2+ through the plasma mebrane. A model is presented suggesting that tentacle retraction is mediated by cytosolic Ca2+ levels which are determined by the fluxing of Ca2+ across the plasma membrane and the membrane of elongate dense bodies which act as internal Ca2+ reservoirs.
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  • 5
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Particle and Particle Systems Characterization 9 (1992), S. 213-222 
    ISSN: 0934-0866
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Modern image analysis equipment has now made it possible to obtain detailed intensity profile information about objects imaged under the optical microscope. If the object contrast is generated by light absorption alone then the image profile of the object is a square wave and the size information is contained in the fundamental frequency of the Fourier components of that square wave. The period of the fundamental frequency lies very close to the intensity midpoint of the image profile. As long as the microscope objective numerical aperture (NA) is high enough to pass this fundamental the object can be sized to an accuracy which is chiefly dependent upon the signal to noise ratio of the system and independent of classical notions of microscope resolution.Thus for latex particle metal replicas it was possible to determine the diameter to a precision which was typically on the order of 13% of the classical Sparrow limit of resolution for the objective employed.By sizing the same particle replica with objectives of different NA it was demonstrated that the size obtained was independent of the objective NA used as long as the replica diameter was above the Sparrow limit. This is in accordance with optical theory.About mid-summer the Goldstein "Zernike" program became available to us through the kindness of Dr. Goldstein. With this program it was possible to model the effect of optical path difference. Unlike particle replicas, most real objects generate object profiles that are a function of refractive index difference and thickness or path difference in addition to object size and transmittance. Although the "Zernike" program can accommodate path differences, it assumes that the object has negligible thickness-an assumption not merited by most real microscopic objects.Although exact quantitative agreement could not always be obtained with reasonable assumptions, the predictions of the "Zernike" program nevertheless could help to define sample preparation conditions which enabled high accuracy sizing to be performed.This work thus demonstrates that an extremely high degree of accuracy and precision in particle sizing is available from the optical microscope which is independent of classical notions of microscope "resolution". The major requirements are that the objective NA be sufficiently high enough to pass the Fourier components which contain the size information and that features in the image can be identified which contain the size information.
    Additional Material: 15 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Particle and Particle Systems Characterization 9 (1992), S. 94-104 
    ISSN: 0934-0866
    Keywords: Chemistry ; Industrial Chemistry and Chemical Engineering
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Of all methods of particle size analysis determination only microscopy and subsequent image analysis enables accurate, geometrically determined size parameters to be measured. For this reason image analysis is a convenient referee or umpire method against which to compare other techniques.In this study, a series of seven poly(vinyl) chloride (PVC) resins were characterized by optical microscopy and image analysis, sieves, HIAC and Coulter counter. It was not found possible to perform the image analysis measurements without operator intervention with a light pen to separate agglomerates. The PVC particles were far too irregular to enable this task to be performed by erosion and dialation techniques. The errors at 90% confidence associated with the image analysis study were: calibration ± 1.7%, counting statistics ± 1% (max) with a focus and algorithms error which ranged from ½-3% depending upon the quantity being measured. These numbers gave a total error of ± 2% (ca. one pixel) for equivalent circular diameter (ECD) and ± 4% (ca. two pixels) for object breadth. In general the agreement between the various techniques was quite good suggesting that the object breadth (OB) as measured by sieves was nearly identical to equivalent spherical diameter (ECD) as measured by the HIAC and Coulter counter for the samples investigates.A study of the image analysis data alone confirmed that OB and ECD were nearly identical with ECD running about 6.1 m̈m above OB.Mass resin samples, which are prone to electrostatic charging effects, tended to give slightly higher results in those methods which measured the resin in the dry state (HIAC and sieves). Finally, it should be suggested that many of the classical concepts of resolution and depth of focus which are used to place limits upon the performance of the optical microscope do not necessarily apply to particle size determination by image analysis. If the chord through the particle which describes its diameter can be considered an approximation to a square wave then it is the fundamental frequency of the Fourier components of that square wave that contains the size information. This should be true for microscopic objects which are absorbing of light and non-retarding in nature.Once this fundamental frequency can be identified then sizing accuracy is dependent chiefly on the signal to noise ratio of the system.
    Additional Material: 13 Ill.
    Type of Medium: Electronic Resource
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