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  • Ca2+ channel blockers Cytoplasmic [Ca2+] transients Fura-2 N-type Ca2+ channels Voltage-gated Ca2+ channels  (1)
  • Signal transduction  (1)
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  • 1
    ISSN: 1432-069X
    Keywords: Key words Keratinocytes ; Intracellular calcium ; Signal transduction ; Heterogeneity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Intracellular calcium release induced by transient applications of phosphoinositide agonists was measured using adherent single HaCaT keratinocytes loaded with the acetoxymethyl derivative of fura-2. Application of ATP, bradykinin and formyl-Met-Leu-Phe (fMLP) resulted in a transient increase in intracellular calcium concentration ([Ca2+]i) with an average half-width of 40 ± 21 s and a decay time constant of 15 ± 10 s (mean ± SD, n = 108), irrespective of the agonist applied. The cells could be classified into two groups: in 53% of the cells repeated stimulation brought about a progressively smaller change in [Ca2+]i (type 1 cells), whereas in the remaining cells the amplitude of the calcium transients was essentially unchanged (type 2 cells). Furthermore, calcium transients in type 1 cells had broader half-widths and slower decays. No difference was found between the agonists in respect of the characteristics of the evoked calcium transient within each subpopulation. However, bradykinin and fMLP desensitized some cells. These results indicate that the activation of the inositol trisphospate transduction pathway by different agonists induces a characteristic elevation of [Ca2+]i within a given cell. Our results demonstrate that cultured HaCaT keratinocytes are heterogeneous in respect of the calcium transients evoked by the activators of this second messenger system.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Ca2+ channel blockers Cytoplasmic [Ca2+] transients Fura-2 N-type Ca2+ channels Voltage-gated Ca2+ channels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract. The significance of voltage-activated Ca2+ currents in eliciting cytoplasmic Ca2+ transients was studied in pyramidal neurones isolated from the rat dorsal cochlear nucleus using combined enzyme treatment/mechanical trituration. Increases in cytoplasmic Ca2+ concentration ([Ca2+]i) were evoked by K+-induced depolarizations (10–50 mM) and monitored by the Fura-2 fluorimetric technique. The acutely dissociated neurones had a resting [Ca2+]i of 17.2±0.5 nM. They possessed caffeine-sensitive Ca2+ stores which were empty at rest; these stores could be filled with Ca2+ entering from the extracellular space and were re-emptied quickly. The effects of various specific high-voltage-activated (HVA) Ca2+ channel antagonists (nifedipine, ω-agatoxin IVA and ω-conotoxin GVIA) on [Ca2+]i transients were tested. Analysis of the blocking effects of these agents on the [Ca2+]i transients indicates that, in the pyramidal neurones of the dorsal cochlear nucleus, N-type Ca2+ channels are primarily responsible for producing the depolarization-induced increases in [Ca2+]i.
    Type of Medium: Electronic Resource
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