Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

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Bahouth, S.W. et al.

Bahouth, Sowinski, Lima,

Regulation of human beta(1)-adrenergic receptors and their mRNA in neuroepithelioma SK-N-MC cells: effects of agonist, forskolin, and protein kinase A.

We determined the effect of long-term exposure to beta-agonists on beta(1)-adrenergic receptors (beta(1)-AR) in human neuroepithelioma SK-N-MC cells because earlier studies have indicated that beta(1)-AR in this cell line are resistant to agonist-induced down-regulation. Exposing SK-N-MC cells to isoproterenol for 24 hr reduced the density of beta(1)-AR by 72%, whereas forskolin, an activator of all the isoforms of adenylyl cyclase, failed to affect the density of beta(1)-AR. Measurement of beta(1)-AR mRNA levels by the ribonuclease protection assay revealed that isoproterenol-induced down-regulation of beta(1)-AR was associated with a sharp decline in beta(1)-AR mRNA, while forskolin also failed to affect this parameter. The differences between the effects of isoproterenol and forskolin on beta(1)-AR were unrelated to cyclic AMP levels, since both agents increased cyclic AMP equally. Next, we determined the role of cyclic AMP-dependent protein kinase A (PKA) in this phenomenon. Inhibition of PKA by its specific inhibitor, H-89 [N-[2-((p-bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide, 2HCl], markedly reduced the magnitude of the isoproterenol-mediated down-regulation of the beta(1)-AR and its mRNA. Transient expression of the catalytic subunit of PKA in SK-N-MC cells down-regulated beta(1)-AR independently of isoproterenol. Therefore, PKA is central to the effect of beta-agonists in down-regulating beta(1)-AR, and its spatial compartmentalization and access to the receptor appear to be essential components of its action.[1]

References

Regulation of human beta(1)-adrenergic receptors and their mRNA in neuroepithelioma SK-N-MC cells: effects of agonist, forskolin, and protein kinase A. Bahouth, S.W., Sowinski, K.M., Lima, J.J. Biochem. Pharmacol. (2001) [Pubmed]

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