Abstract
The purpose of the present study was to determine the molecular mechanism of stimulatory actions of ACh and vasoactive intestinal polypeptide (VIP) by determining the role of various second messengers in the neurohumoral secretion. Toward such a goal, we measured cAMP, cGMP, protein kinase (PKC) activity, 3H-inositol triphosphate (3H-IP3), and 45Ca uptake in the adrenal medulla subjected to various treatments. Stimulation of splanchnic nerve endings increased 45Ca uptake, cAMP content, 3H-IP3, and PKC activity in the adrenal medulla. If muscarinic receptors of chromaffin cells were selectively activated by perfusion with muscarine, 3H-IP3 content and PKC activity were enhanced. Nicotine, on the other hand, increased only 45Ca uptake without affecting any other second messenger. Perfusion with VIP increased PKC activity and cAMP and 3H-IP3 content. None of the procedures affected cGMP content. Interplay among various second messengers was further investigated by studying interactions of nicotinic, muscarinic, and VIP- ergic receptors in modulation of catecholamine (CA) secretion and by using agents known to activate specific second messengers (e.g., forskolin, phorbol esters). Our results show that muscarine, VIP, and phorbol ester facilitated nicotine-evoked secretion by increasing PKC activity, and it was associated with an additional increase in 45Ca accumulation. On the other hand, secretion evoked by nicotine as well as muscarine was facilitated by forskolin without additional increase in 45Ca accumulation. A novel feature of the study is that ACh and VIP activate three types of receptors on chromaffin cells to stimulate and mutually facilitate the secretion of CA by generating various second messengers.(ABSTRACT TRUNCATED AT 250 WORDS)