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

Boyano-Adanez, M C; Lundqvist, C; Larsson, Christer LU and Gustavsson, L (1997) In Neuropharmacology 36(3). p.295-304
Abstract
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
G-protein, acetylcholine, muscarinic receptor, phosphatidylethanol, phospholipase D, protein kinases, signal transduction
in
Neuropharmacology
volume
36
issue
3
pages
295 - 304
publisher
Elsevier
external identifiers
  • pmid:9175607
  • scopus:0343416940
ISSN
1873-7064
DOI
10.1016/S0028-3908(96)00178-5
language
English
LU publication?
yes
id
1b8edece-ac08-4e30-9e8c-455b71ce3ca9 (old id 1111182)
date added to LUP
2008-07-16 16:54:33
date last changed
2017-01-01 04:37:55
@article{1b8edece-ac08-4e30-9e8c-455b71ce3ca9,
  abstract     = {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.},
  author       = {Boyano-Adanez, M C and Lundqvist, C and Larsson, Christer and Gustavsson, L},
  issn         = {1873-7064},
  keyword      = {G-protein,acetylcholine,muscarinic receptor,phosphatidylethanol,phospholipase D,protein kinases,signal transduction},
  language     = {eng},
  number       = {3},
  pages        = {295--304},
  publisher    = {Elsevier},
  series       = {Neuropharmacology},
  title        = {Characterization of phospholipase D activation by muscarinic receptors in human neuroblastoma SH-SY5Y cells},
  url          = {http://dx.doi.org/10.1016/S0028-3908(96)00178-5},
  volume       = {36},
  year         = {1997},
}