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Muscarinic-receptor-mediated changes in intracellular Ca2+ and inositol 1,4,5-trisphosphate mass in a human neuroblastoma cell line, SH-SY5Y.

Abstract
This study reports increased intracellular Ca2+ and inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] in response to muscarinic-cholinergic stimulation of human neuroblastoma (SH-SY5Y) cells. Carbachol stimulation leads to a rapid increase in intracellular Ca2+ and Ins(1,4,5)P3 mass, both reaching a peak at around 10 s and then declining to a new maintained phase significantly above basal. Dose-response analysis of peak and plateau phases of intracellular Ca2+ shows different agonist potencies for both phases, carbachol being more potent for the plateau phase. The plateau-phase intracellular Ca2+ was dependent on extracellular Ca2+, which is admitted to the cell through a non-voltage-sensitive Ni2(+)-blockable Ca2+ channel. Using a Mn2+ quench protocol, we have shown that Ca2+ entry occurs early during the discharge of the internal stores. The plateau phase (Ca2(+)-channel opening) is dependent on the continued presence of agonist, since addition of atropine closes the Ca2+ channel and intracellular Ca2+ declines rapidly back to basal. We also failed to detect a refilling transient when we added back Ca2+ after intracellular Ca2+ had reached a peak and then declined in Ca2(+)-free conditions. These data strongly suggest that muscarinic stimulation of SH-SY5Y cells leads to a rapid release of Ca2+ from an Ins(1,4,5)P3-sensitive internal store and a parallel early entry of Ca2+ across the plasma membrane.
AuthorsD G Lambert, S R Nahorski
JournalThe Biochemical journal (Biochem J) Vol. 265 Issue 2 Pg. 555-62 (Jan 15 1990) ISSN: 0264-6021 [Print] England
PMID2302186 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Benzofurans
  • Fluorescent Dyes
  • Receptors, Muscarinic
  • Manganese
  • Arecoline
  • Ionomycin
  • Atropine
  • Muscarine
  • Inositol 1,4,5-Trisphosphate
  • Carbachol
  • Calcium
  • Fura-2
Topics
  • Arecoline (pharmacology)
  • Atropine (pharmacology)
  • Benzofurans
  • Calcium (metabolism)
  • Carbachol (pharmacology)
  • Cell Line
  • Fluorescent Dyes
  • Fura-2
  • Humans
  • Inositol 1,4,5-Trisphosphate (metabolism)
  • Ionomycin (pharmacology)
  • Kinetics
  • Manganese (pharmacology)
  • Muscarine (pharmacology)
  • Neuroblastoma
  • Receptors, Muscarinic (drug effects, physiology)
  • Tumor Cells, Cultured (drug effects, metabolism)

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