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Release and sequestration of Ca2+ by a caffeine- and ryanodine-sensitive store in a sub-population of human SH-SY5Y neuroblastoma cells

Lookup NU author(s): Fiona Riddoch, Sophie Rowbotham, Dr Anna Brown, Dr Chris Redfern, Dr Timothy Cheek

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Abstract

We have used single cell fluorescence imaging techniques to examine the role that ryanodine receptors play in the stimulus-induced Ca2+ responses of SH-SY5Y cells. The muscarinic agonist methacholine (1 mM) resulted in a Ca2+ signal in 95% of all cells. Caffeine (30 mM) however stimulated a Ca2+ signal in only 1-7% of N-type (neuroblastic) cells within any given field. The caffeine response was independent of extracellular Ca2+, regenerative in nature, and abolished in a use-dependent fashion by ryanodine. In caffeine-responsive cells, the magnitude of the methacholine-induced Ca2+ signal was inhibited by 75.07 ± 5.51% by pretreatment with caffeine and ryanodine, suggesting that the caffeine-sensitive store may act as a Ca2+ source after muscarinic stimulation. When these data were combined with equivalent data from non-caffeine-responsive cells, the degree of apparent inhibition was significantly reduced. In contrast, after store depletion by caffeine, the Ca2+ signal induced by 55 mM K+ was potentiated 2.5-fold in the presence of ryanodine, suggesting that the store may act a Ca2+ sink after depolarisation. We conclude that a caffeine- and ryanodine-sensitive store can act as a Ca2+ source and sink in SH-SY5Y cells, and that effects of the store can become obscured if data from caffeine-insensitive cells are not excluded. © 2005 2005 Elsevier Ltd. All rights reserved.


Publication metadata

Author(s): Riddoch, F.C., Rowbotham, S.E., Brown, A.M., Redfern, C.P.F., Cheek, T.R.

Publication type: Article

Publication status: Published

Journal: Cell Calcium

Year: 2005

Volume: 38

Issue: 2

Pages: 111-120

Print publication date: 01/08/2005

ISSN (print): 0143-4160

ISSN (electronic): 1532-1991

URL: http://dx.doi.org/10.1016/j.ceca.2005.06.001

DOI: 10.1016/j.ceca.2005.06.001

PubMed id: 16095688


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