Muscarinic regulation of Ca2+ oscillation frequency in GH3 cells

The GH3 anterior pituitary cell line has been used as a model to investigate diverse aspects of pituitary cell physiology including Ca2+ homeostasis and secretion. These cells possess muscarinic receptors which, by activating K+ channels and inhibiting Ca2+ channels, should decrease electrical excitability. We measured the effect of carbachol (10 microM) on the frequency of Ca2+ oscillations caused by Ca2+ action potentials in the plasma membrane. Carbachol reduced oscillation frequency by approximately 85% (p < 0.001). This inhibition was reversed by atropine (1 microM), and was prevented by pre-incubation with pertussis toxin (200 ng/ml, 24 h). Since many anterior pituitary cell types secrete acetylcholine, the presence of muscarinic receptors coupled to cell excitability in these cells suggest that ACh could exert a paracrine- or autocrine-like action in GH3 cell cultures. In experiments designed to test this idea, perfusion with 1 microM atropine caused a small but significant increase (p < 0.05) in oscillation frequency when the cells had previously been incubated for 30 min without perfusion. However, this effect was not blocked by either pre-treatment with pertussis toxin or by including atropine during the entire experiment (including the 30-min incubation without perfusion). We conclude that these cells respond to muscarinic agonists by decreasing oscillation frequency but find no evidence for feedback control by endogenous ACh under these conditions.