Characterization of phospholipase D activation by muscarinic receptors in human neuroblastoma SH-SY5Y cells

M C Boyano-Adanez, C Lundqvist, Christer Larsson, L Gustavsson

Research output: Contribution to journalArticlepeer-review


The cholinergic regulation of phospholipase D activity was studied in SH-SY5Y human neuroblastoma cells with phosphatidylethanol formation as a specific marker for the enzyme activity. The muscarinic antagonists, hexahydrosiladifenidol and pirenzepine, inhibited carbachol-induced phosphatidylethanol formation in a concentration-dependent manner and the inhibitory constants indicated that muscarinic M1 receptors are responsible for the major part of the phospholipase D activation. The mechanism of receptor-mediated phospholipase D activation varies between different cell types and receptors. In SH-SY5Y cells, the carbachol-induced phospholipase D activity was inhibited by protein kinase C inhibitors. Since both phospholipases D and C are activated by muscarinic stimulation in SH-SY5Y cells, most of the phospholipase D activation is probably secondary to the protein kinase C activation that follows phospholipase C-mediated increase in diacylglycerols. Other kinases may be involved in the regulation since also a tyrosine kinase inhibitor decreased the phosphatidylethanol formation. Stimulation of G-protein(s) and increase in the intracellular Ca2+ concentration activated phospholipase D and may be additional mechanisms for the muscarinic regulation of phospholipase D in SH-SY5Y cells. Propranolol, an inhibitor of phosphatidic acid phosphohydrolase, increased the carbachol-induced formation of phosphatidic acid at the expense of 1,2-diacylglycerol. This indicates that phospholipase D contributes to the formation of 1,2-diacylglycerol after carbachol stimulation in SH-SY5Y cells.
Original languageEnglish
Pages (from-to)295-304
Issue number3
Publication statusPublished - 1997

Bibliographical note

The information about affiliations in this record was updated in December 2015.
The record was previously connected to the following departments: Tumour Cell Biology (013017530)

Subject classification (UKÄ)

  • Pharmacology and Toxicology


  • G-protein
  • acetylcholine
  • muscarinic receptor
  • phosphatidylethanol
  • phospholipase D
  • protein kinases
  • signal transduction


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