Involvement of the muscarinic acetylcholine receptor in inhibition of cell migration. Biochem Pharmacol 2002 Feb 15;63(4):597-605
Date
05/07/2002Pubmed ID
11992627DOI
10.1016/s0006-2952(01)00901-7Scopus ID
2-s2.0-0037085006 (requires institutional sign-in at Scopus site) 8 CitationsAbstract
Activation of G protein-coupled receptors is known to stimulate cell migration, but receptor-mediated signals inhibiting cell migration have not been identified. We investigated the ability of transfected human M(3) muscarinic acetylcholine receptors (mAChR) to regulate the migration of Chinese hamster ovary (CHO) cells. Single cells migrated on colloidal gold applied to fibronectin-coated plates, and videomicroscopy was used to measure cell spreading and migration. Activation of M(3) mAChR with the agonist carbachol was found to inhibit cell migration, whereas direct activation of protein kinase C (PKC) with PMA was found to stimulate migration. The amount of cell adhesion and spreading was found to be equivalent for carbachol- and PMA-treated cells. Selective inactivation of conventional PKC isoforms with Go6976 (C(24)H(18)N(4)O) abolished the PMA-mediated increase in cell migration. In contrast, the mAChR-mediated decrease in migration was not altered by Go6976, but was abolished when both novel and conventional PKC isoforms were inactivated by calphostin C or chelerythrine. These findings suggest involvement of conventional PKC isoforms in the stimulation of migration and of novel PKC isoforms in the inhibition of migration. Carbachol- but not PMA-treated cells exhibited an elongated morphology reminiscent of migrating cells that cannot detach their trailing edges from the substratum. Similarly, carbachol-treated cells detached less readily from fibronectin than control or PMA-treated cells when integrin activity was diminished by the chelation of Ca(2+) and Mg(2+). Finally, the carbachol-induced diminution of cell detachment was preserved after inhibition of the conventional PKC isoforms with Go6976, but was abrogated by treatment with either calphostin C or chelerythrine. These findings suggest that mAChR activation diminishes the ability of cells to detach from the substratum, resulting in diminished migration. This is in contrast to the direct activation of PKC with PMA, which stimulates migration.
Author List
Varker KA, Williams CLAuthor
Carol L. Williams PhD Professor in the Pharmacology and Toxicology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsCHO Cells
Carbachol
Cell Adhesion
Cell Movement
Cholinergic Agonists
Cricetinae
Dose-Response Relationship, Drug
Fibronectins
Humans
Isoenzymes
Protein Kinase C
Receptors, Muscarinic
Tetradecanoylphorbol Acetate
Transfection