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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Excitation-contraction coupling ; Cell calcium ; Mammalian cardiac muscles ; Reversibly hyperpermeable cells ; Calcium indicators ; Aequorin ; Isoproterenol ; Caffeine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The bioluminescent calcium indicator aequorin was successfully loaded into mammalian working myocardium of ferrets by a chemical procedure which makes the cells reversibly hyperpermeable through exposure to Ethylenebis-(oxyethylenenitrilo) tetraacetic acid (EGTA). After undergoing the loading procedure, developed tension at Lmax was 103±26% of the control, which indicates that the muscles regained normal function. The configurations of the aequorin signals (i.e., calcium transients) and their responses to drugs were the same as reported after microinjection of aequorin. The peak of the Ca++ transient determined by the method of fractional luminescence at 3s intervals of stimulation, 2.5mM [Ca++]o, 22.5°C was 1.1μM; this value is similar to that reported for microinjection. These results indicate that the chemical loading procedure is a useful alternative to microinjection for loading aequorin into mammalian working myocardium.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2013
    Keywords: Quin 2 ; Aequorin ; Vascular Smooth Muscle ; Mammalian Isolated Cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A comparison between the fluorescent indicator quin 2 and the bioluminescent indicator aequorin was performed in the same smooth muscle cell type. Aequorin was loaded into intact strips and quin was loaded into enzymatically isolated single cells from ferret portal vein. Both indicators gave qualitatively the same calcium profiles when the tissue was challenged with agonists. Quin loading caused a dramatic shift to the right in dose response curves to potassium and phenylephrine. The ED50 values for quin loaded cells were significantly different from those for control cells for both agonists. Intracellular calcium levels at rest were not significantly different with quin and aequorin. Cells stimulated with potassium gave significantly different intracellular calcium values with the two indicators suggesting a change in the stimulated steady state level due to the introduction of an additional calcium buffer (quin2) into the cell.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2013
    Keywords: Mammalian vascular smooth muscle ; Enzymatically isolated cells ; Vasodilators
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A modified method for enzymatically isolating mammalian vascular smooth muscle cells has been developed and tested for ferret portal vein smooth muscle. This method produces a high proportion of fully relaxed cells and these cells appear to have normal pharmacological responsiveness. The ED50 values for both alpha stimulation and potassium depolarization are not significantly different in the isolated cells from those obtained from intact strips of ferret portal vein, suggesting that the enzymatic treatment does not destroy receptors or alter the electrical responsiveness of the cells. It was also possible to demonstrate a vasodilatory action of papaverine, nitroprusside and adenosine directly on the isolated cells indicating that the pathways involved are intact in the isolated cells. This method should be of considerable usefulness, particularly in combination with the new fluorescent indicators and cell sorter techniques which require isolated cells.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2013
    Keywords: Smooth Muscle ; Calcium ; PDGF ; Aequorin ; Angiotensin ; Atherosclerosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We studied stimulus-specific alterations of the excitation-contraction coupling pathway in freshly isolated contractile and subcultured non-contractile vascular smooth muscle cells. Using the calcium indicator aequorin, we detected physiological increases in cytoplasmic free calcium ([Ca2+]i) in subcultured smooth muscle cells subjected to angiotensin or 33 mM potassium depolarization. These increases were qualitatively identical to those previously measured in intact vascular strips. Platelet-derived growth factor (PDGF) induced a slow, sustained [Ca2+]i increase when applied to the subcultured smooth muscle cells at low picomolar concentrations. Freshly isolated, contractile vascular smooth muscle cells, prepared by a novel technique, exhibited a slow shortening of 20% of resting length in response to PDGF. PDGF also markedly potentiated smooth muscle cell shortening in response to an ED50 dose of phenylephrine. This effect was PDGF concentration dependent. The time course of shortening induced by PDGF alone was consistent with the time course of the PDGF-induced [Ca2+]i increase in the cultured smooth muscle cells. These data suggest that agonists which induce [Ca2+]i changes in contractile smooth muscle cells may retain this ability with respect to cultured smooth muscle cells. PDGF, a peptide mitogen for proliferative smooth muscle cells, may also serve to modulate vascular tone by modestly raising [Ca2+]i in contractile smooth muscle cell and, therefore, sensitizing the cells to alpha adrenergic agonists.
    Type of Medium: Electronic Resource
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