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  • 1
    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.
    Type of Medium: Electronic Resource
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  • 2
    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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  • 3
    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.
    Type of Medium: Electronic Resource
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