Interaction between Adrenergic Agonists and Forskolin on Adenylate Cyclase Activity in the Rabbit Proximal Tubule

Abstract

Using a micro-radioimmunoassay, cAMP was measured in single, isolated S2 proximal straight tubules dissected from rabbit kidneys to investigate the effects of adrenergic agonists on adenylate cyclase activity. The baseline activity of adenylate cyclase was low and unaffected by either the α₂ agonist clonidine, the β agonist isoprenaline (in the absence or presence of 1 µM forskolin) or 1 µM forskolin. Adenylate cyclase activity was markedly stimulated by 20 µM forskolin, an effect which was inhibited by 1 µM clonidine. The inhibition by clonidine was not apparent in the presence of the α₂ antagonist yohimbine. These results confirm the inability of isoprenaline to stimulate adenylate cyclase in the rabbit proximal tubule and demonstrate the coupling of α₂ receptors, in an inhibitory fashion, to adenylate cyclase. The inhibitory action of the α₂-receptor agonist was independent of other hormone activity in the renal proximal tubule.

Minute-scale persistence of a GPCR conformation state triggered by non-cognate G protein interactions primes signaling

“…Stimulation was performed with 0.3 nM to 100 μM fenoterol in the presence of 2 μM forskolin for 10 min. As the response was being monitored from endogenously expressed β 2 -AR, it had a low magnitude and was synergized by the addition of forskolin 61 . The concentration of forskolin used did not produce a significant response in the absence of fenoterol (Supplementary Fig.…”