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  • 1
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Cell Differentiation 23 (1988), S. 77-86 
    ISSN: 0045-6039
    Keywords: Brain ; Ca^2^+ mobilization ; Chick embryo ; Heart ; Morphogenesis ; Muscarinic receptor
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-041X
    Keywords: Calcium mobilization ; Muscarinic cholinergic receptor ; Chlorotetracycline ; Morphogenesis ; Limb bud
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cell suspensions of chick limb buds (stage 23/24) were loaded with the fluorescent Ca2+ chelator chlorotetracycline. Fluorescence was monitored in a spectrofluorometer. Stimulation with acetylcholine induced a fluorescence decrease, indicating intracellular Ca2+ mobilization. The fluorescence decrease triggered by acetylcholine was inhibited by muscarinic but not by nicotinic antagonists, indicating that a muscarinic acetylcholine receptor is involved. The muscarinic receptor in the chick limb bud has a characteristic pharmacological profile: acetylcholine, carbachol and acetyl-β-methylcholine functioned as full agonists triggering maximal fluorescence decrease. Bethanechol was less effective, producing only one-third of the maximum response. Pilocarpine and oxotremorine, two classical agonists in other systems, were ineffective and functioned as antagonists. In the chick limb bud, cholinesterase, choline acetyltransferase and the presence of a muscarinic receptor have been demonstrated in previous studies. The present experiments show that stimulation of the embryonic muscarinic receptor leads to intracellular Ca2+ mobilization.
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  • 3
    ISSN: 1432-041X
    Keywords: Muscarinic receptors ; Acetylcholinersterase ; Morphogenesis ; Ecdysone ; Chironomus cell line
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The permanent epithelial insect cell line used was derived from Chironomus tentans embryos. Cells are maintained in suspension culture, where they grow as single-layered vesicles. On treatment with the moulting hormone 20-OH-ecdysone cell division ceases. Patches of cuboidal epithelium appear in the vesicles which finally become multilayered and form bud-like protrusions at the outside. In the present study, we localized cholinesterase activity in the cell protrusions by histochemistry and demonstrated coexpression of cholinergic muscarinic receptors by immunofluorescence. Muscarinic receptors were visualized with the monoclonal antibody M35. Six hours after treatment with 20-OH-ecdysone, muscarinic receptors appeared in a few individual cells of the epithelial vesicles before morphological changes became visible. After 24 h, immunofluorescence was concentrated in multilayered patches which now also showed cholinesterase activity. After 3 days, muscarinic receptors and cholinesterase activity were localized in the epithelium protrusions. The results are discussed in the context of an embryonic cholinergic muscarinic system the expression of which has been described in vertebrate and non-vertebrate embryos and is correlated with phases of morphogenesis.
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  • 4
    ISSN: 1432-041X
    Keywords: Phorbol ester ; Muscarinic receptor ; Calcium ; Chick embryo ; Morphogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A muscarinic cholinergic receptor is present on undifferentiated cells of the chick embryo. Stimulation of the muscarinic receptor with muscarinic agonists triggers intracellular Ca2#x002B; mobilization. Here, we investigate the effect of phorbol 12-myristate 13-acetate (PMA) on the muscarinic receptor-mediated Ca2#x002B; mobilization, which is monitored in cell suspensions of chick embryos of stage 24 by chlorotetracycline fluorescence. PMA inhibits the Ca2#x002B; mobilization in a time-dependent and concentration-dependent manner without changing the ED50 of acetylcholine. The concentration of PMA that gives halfmaximal inhibition is 3.1×10−9 M PMA.
    Type of Medium: Electronic Resource
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