Ca2+ entry following store depletion in SH-SY5Y neuroblastoma cells.

Abstract

Ca2+ entry following Ca2+ store depletion was examined in the human neuroblastoma cell line, SH-SY5Y, by measuring the concentration of intracellular free Ca2+ ([Ca2+]i) with fura-2. Application of the muscarinic agonist oxotremorine-M (oxo-M) caused an increase in [Ca2+]i. This consisted of a peak, mediated by release of Ca2+ from internal stores followed by a sustained plateau, mediated by Ca2+ entry across the plasma membrane. The Ca2+ entry resulted from depletion of intracellular Ca2+ stores This pathway was further characterized in the presence of thapsigargin, an inhibitor of the Ca2+ ATPase involved in replenishing IP3-sensitive stores. Stores were first depleted with oxo-M and thapsigargin in the absence of extracellular Ca2+. After washout of oxo-M, subsequent exposure to Ca2+ evoked reproducible increases in [Ca2+]i. Application of oxo-M plus Ca2+ had little effect on the increases in [Ca2+]i, indicating that in SH-SY5Y cells, agonist-dependent pathways contribute little to Ca2+ entry following store depletion. Mn2+, Sr2+ and Ba2+ were permeable through this pathway. Mn2+ and Ba2+ also showed slight permeability in the absence of store depletion. Ca2+ entry following store depletion was blocked by La3+ (IC50 = 75 nM) and by SKF 96365. La3+ blocked Mn2+ entry through the pathway activated by store depletion but did not affect basal Mn2+ permeability. These results indicate that SH-SY5Y neuroblastoma cells have an agonist-independent Ca2+ entry pathway activated by store depletion.

• Authors: T J Grudt, M M Usowicz, G Henderson
• Journal: Brain research. Molecular brain research (Brain Res Mol Brain Res) Vol. 36 Issue 1 Pg. 93-100 (Feb 1996) ISSN: 0169-328X [Print] Netherlands
• PMID: 9011769 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Imidazoles
• Muscarinic Agonists
• Barium
• Manganese
• Oxotremorine
• oxotremorine M
• Thapsigargin
• Lanthanum
• Inositol 1,4,5-Trisphosphate
• 1-(2-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenylethyl)-1H-imidazole
• Calcium
• Strontium

Topics

• Barium (pharmacokinetics)
• Calcium (metabolism)
• Cell Membrane Permeability
• Imidazoles (pharmacology)
• Inositol 1,4,5-Trisphosphate (metabolism)
• Lanthanum (pharmacology)
• Manganese (pharmacokinetics)
• Muscarinic Agonists (pharmacology)
• Neuroblastoma
• Neurons (metabolism)
• Oxotremorine (analogs & derivatives, pharmacology)
• Strontium (pharmacokinetics)
• Thapsigargin (pharmacology)
• Tumor Cells, Cultured