Capacitative calcium entry supports calcium oscillations in human embryonic kidney cells.
Treatment of human epithelial kidney (HEK293) cells with low concentrations of the muscarinic agonist methacholine results in the activation of complex and repetitive cycling of intracellular calcium ([Ca(2+)](i)), known as [Ca(2+)](i) oscillations. These oscillations occur with a frequency that depends on the concentration of methacholine, whereas the magnitude of the [Ca(2+)](i) spikes does not. The oscillations do not persist in the absence of extracellular Ca(2+), leading to the conclusion that entry of Ca(2+) across the plasma membrane plays a significant role in either their initiation or maintenance. However, treatment of cells with high concentrations of GdCl(3), a condition which limits the flux of calcium ions across the plasma membrane in both directions, allows sustained [Ca(2+)](i) oscillations to occur. This suggests that the mechanisms that both initiate and regenerate [Ca(2+)](i) oscillations are intrinsic to the intracellular milieu and do not require entry of extracellular Ca(2+). This would additionally suggest that, under normal conditions, the role of calcium entry is to sustain [Ca(2+)](i) oscillations. By utilizing relatively specific pharmacological manoeuvres we provide evidence that the Ca(2+) entry that supports Ca(2+) oscillations occurs through the store-operated or capacitative calcium entry pathway. However, by artificial introduction of a non-store-operated pathway into the cells (TRPC3 channels), we find that other Ca(2+) entry mechanisms can influence oscillation frequency in addition to the store-operated channels.
Bird
Gary St J
GS
National Institute of Environmental Health Sciences/NIH, PO Box 12233, Research Triangle Park, NC 27709, USA. bird@niehs.nih.gov
Putney
James W
JW
D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000187:drug effects / Q000502:physiology*; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D025802:Electric Capacitance; D005682:Gadolinium / Q000008:administration & dosage*; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology*; D007668:Kidney / Q000187:drug effects / Q000196:embryology / Q000502:physiology*; D007700:Kinetics; D008657:Metabolic Clearance Rate; D016210:Methacholine Chloride / Q000008:administration & dosage*
D015220:Calcium Channels; D016210:Methacholine Chloride; D005682:Gadolinium; C038958:gadolinium chloride; D002118:Calcium