Regulation of muscarinic M2 receptors.
The molecular mechanisms involved in the regulation of muscarinic receptor gene expression are poorly understood. In an effort to gain a better understanding of the regulation of M2 receptors, we have investigated homologous and heterologous regulation of M2 muscarinic receptor protein and gene expression in human embryonic lung fibroblasts (HEL 299 cells). HEL 299 cells constitutively express m2 receptors, with no evidence of other muscarinic receptor subtypes. We have shown that M2 receptors in these cells can be down-regulated by muscarinic and beta2-adrenergic receptor agonists. Unlike the down-regulation mediated by muscarinic and beta-adrenergic stimulation, activation of PKC with PDBu was mediated through changes in m2 muscarinic receptor mRNA through reduced gene transcription. Because of the inflammatory nature of asthma, we have focused on delineating the interactions between cytokines and M2 receptors in an attempt to define potential endogenous modulators of M2 receptor expression. We have shown that the multi-functional cytokine, transforming growth factor beta1 (TGF-beta1), which is involved in several inflammatory conditions induces desensitization and down-regulation of M2 muscarinic receptor protein and gene expression that was mediated through a reduction in the rate of m2 receptor gene transcription. Other cytokines of interest are tumor necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL-1beta) which are elevated in asthma. We have demonstrated that TNF-alpha and IL-1beta synergise to induce down-regulation of M2 muscarinic receptor protein and mRNA which was associated with functional desensitization of the receptor protein. The M2 receptor mRNA down-regulation appeared to be mediated through a reduction in the rate of m2 receptor gene transcription which may be dependent on the transcription and translation of unknown protein factor(s). Moreover, a role of PKA and ceramide pathways in M2 receptor regulation is suggested. Collectively, our work provides a mechanistic explanation of previous reports indicating altered function of M2 receptors in asthma. Ours results also suggest that the expression of this receptor subtype may be under the control of a cytokine network at the airways.
Barnes
P J
PJ
Department of Thoracic Medicine, Imperial College School of Medicine at National Heart and Lung Institute, London, UK.
Haddad
E B
EB
Rousell
J
J
D000818:Animals; D004789:Enzyme Activation; D006801:Humans; D018721:Muscarinic Agonists / Q000494:pharmacology; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D043585:Receptor, Muscarinic M2; D018343:Receptors, Adrenergic, beta-2 / Q000502:physiology; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000502:physiology*; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D018721:Muscarinic Agonists; D010982:Platelet-Derived Growth Factor; D043585:Receptor, Muscarinic M2; D018343:Receptors, Adrenergic, beta-2; D011976:Receptors, Muscarinic; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D011493:Protein Kinase C