Differential agonist-induced regulation of human M2 and M 3 muscarinic receptors

Matthias B. Stope, Christina Kunkel, Christian Kories, Martina Schmidt, Martin C. Michel*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

9 Citations (Scopus)

Abstract

We have compared the regulation of M2 and M3 muscarinic receptors heterologously expressed in HEK-293 cells upon long-term exposure towards the agonist carbachol. Carbachol time- and concentration- dependently reduced M2 receptor density with a maximum reduction of about 60%. Treatment with 1mM carbachol for 24hr was accompanied by desensitisation of carbachol-induced Ca2+ elevations (maximum response reduced by 70%) but not by alterations in the expression of various G-protein α-subunits. Consistently, heterologous desensitisation of Ca2+ elevations by the purinergic receptor agonist ATP or by sphingosine-1-phosphate was not detected. In contrast, carbachol time- and concentration-dependently up-regulated M3 receptors with maximum increases to about 350% of control values. The up-regulation was fully blocked by cycloheximide indicating that it was dependent on protein synthesis. Concomitant with the up-regulation of the M3 receptor was a reduction in the expression of the α-subunit of Gq/11. The net effect of these two opposite regulatory mechanisms was a lack of alteration of carbachol-stimulated Ca2+ elevation. However, the reduction of G q/11 was accompanied by a heterologous desensitisation of Ca 2+ elevations by ATP and sphingosine-1-phosphate. Levels of M 2 and M3 receptor mRNA as assessed by real-time PCR were not significantly altered by carbachol exposure for either receptor, suggesting that alterations of mRNA stability did not contribute to the observed changes in receptor number. We conclude that M2 and M3 receptor expression within the same cell undergoes differential agonist-induced regulation being accompanied by distinct regulation of G-protein expression leading to differential effects on signal transduction by other receptor systems.

Original languageEnglish
Pages (from-to)2099-2105
Number of pages7
JournalBiochemical Pharmacology
Volume66
Issue number11
DOIs
Publication statusPublished - 1-Dec-2003
Externally publishedYes

Keywords

  • ATP
  • Calcium
  • G-protein
  • Muscarinic receptor
  • Regulation
  • Sphingosine-1- phosphate

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