title: Spontaneous Ca2+ spiking in a vascular smooth muscle cell line is independent of the release of intracellular Ca2+ stores.
abstract: Monolayers of fura-2-loaded A7r5 cells, a cell line derived from rat embryonic aorta, generated spontaneous Ca2+ spikes that were synchronized within the cell population. These Ca2+ spikes were abolished by removal of extracellular Ca2+ or addition of nimodipine (50 nM), and their frequency was increased by depolarization with high K+ or by treatment with BAYK 8644 (1 microM), indicating that Ca2+ entry through L-type Ca2+ channels is required for Ca2+ spiking. Several lines of evidence indicate that mobilization of intracellular Ca2+ stores is not necessary for this Ca2+ spiking. 1) Ryanodine (0.1-50 microM) neither stimulated Ca2+ mobilization nor affected Ca2+ spiking; 2) the complex effects of caffeine were mimicked by theophylline, 8-bromo-cyclic adenosine 3':5'-monophosphate (8-bromo-cAMP), and forskolin, suggesting that the caffeine effects may be mediated by cAMP and not by ryanodine receptors; 3) prolonged incubation with thapsigargin (50 nM), which depletes intracellular Ca2+ stores, did not affect the frequency of Ca2+ spiking; 4) Ba2+ or Sr2+ could substitute for Ca2+ in the spike-generating mechanism even when intracellular stores were depleted of Ca2+. Under conditions where the sarcoplasmic reticulum (SR) contained Ca2+, Ba2+ spikes did not cause Ca2+ mobilization. The mechanisms involved in generating spontaneous Ca2+ spiking in A7r5 cells are therefore likely to reside in the sarcolemma and to operate independently of SR Ca2+ uptake and release.
authors: Department of Pharmacology, University of Cambridge, United Kingdom.
authors: K L
authors: KL
authors: Byron
authors: C W
authors: CW
authors: Taylor
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D004563:Electrochemistry; D016257:Fura-2; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D009553:Nimodipine / Q000494:pharmacology; D051381:Rats; D013050:Spectrometry, Fluorescence
chemical_list: D009553:Nimodipine; D002118:Calcium; D016257:Fura-2