M3-subtype muscarinic receptor activation stimulates intracellular calcium oscillations and aldosterone production in human adrenocortical HAC15 cells

Abstract

A previous body of work in bovine and rodent models shows that cholinergic agonists modulate the secretion of steroid hormones from the adrenal cortex. In this study we used live-cell Ca²⁺ imaging to investigate cholinergic activity in the HAC15 human adrenocortical carcinoma cell line. The cholinergic agonists carbachol and acetylcholine triggered heterogeneous Ca²⁺ oscillations that were strongly inhibited by antagonists with high affinity for the M₃ muscarinic receptor subtype, while preferential block of M₁ or M₂ receptors was less effective. Acute exposure to carbachol and acetylcholine modestly elevated aldosterone secretion in HAC15 cells, and this effect was also diminished by M₃ inhibition. HAC15 cells expressed relatively high levels of mRNA for M₃ and M₂ receptors, while M₁ and M₅ mRNA were much lower. In conclusion, our data extend previous findings in non-human systems to implicate the M₃ receptor as the dominant muscarinic receptor in the human adrenal cortex.

Keywords: CHRM2; CHRM3; Fluo-4; Hypertension; Methoctramine; Tiotropium.