Genetic Profiling of α1-Adrenergic Receptor Subtypes by Oligonucleotide Microarrays: Coupling to Interleukin-6 Secretion but Differences in STAT3 Phosphorylation and gp-130

Adrenoceptor subtypes (1A-, 1B-, 1D-) are known to cou- ple to similar signaling pathways, although differences among the subtypes do exist. As a more sensitive assay, we used oligonucleotide microarrays to identify gene expression changes in Rat-1 fibroblasts stably expressing each individual subtype. We report the gene expressions that change by at least a factor of 2 or more. Gene expression profiles signifi- cantly changed equally among all three subtypes, despite the unequal efficacy of the inositol phosphate response. Gene ex- pressions were clustered into cytokines/growth factors, tran- scription factors, enzymes, and extracellular matrix proteins. There were also a number of individual subtype-specific changes in gene expression, suggesting a link to independent pathways. In addition, all three 1-AR subtypes robustly stim- ulated the transcription of the prohypertrophic cytokine inter- leukin (IL)-6, but differentially altered members of the IL-6 sig- naling pathway (gp-130 and STAT3). This was confirmed by measurement of secreted IL-6, activated STAT3, and gp-130 levels. Activation of STAT3 Tyr705 phosphorylation by the 1- ARs was not through IL-6 activation but was synergistic with IL-6, suggesting direct effects. Interestingly, 1B-AR stimulation caused the dimerization-dependent phosphorylation of Tyr705 on STAT3 but did not activate the transcriptional-dependent phosphorylation of Ser727. The 1B-AR also constitutively down-regulated the protein levels of gp-130. These results suggest that the 1B-AR has differential effects on the phos- phorylation status of the STAT3 pathway and may not be as prohypertrophic as the other two subtypes.