FURTHER CHARACTERIZATION OF 5-HT-RECEPTOR AND 5-HT(3)-RECEPTOR AGONISTS-STIMULATED PHOSPHOINOSITOL PHOSPHATES ACCUMULATION

The calcium requirement for serotonin (5-HT)- and the 5-HT3 receptor agonists, 2-Me-5-HT- and PBG-dependent breakdown of phosphatidyl inositol has been examined in the rat fronto-cingulate cortex. The omission of added Ca2+ from the Kreb's incubation medium reduced the [H-3]inositol phosphate accumulation from pre-labelled phospholipids. Removal of Ca2+ by pre-incubation with EGTA (0.5 mM), as well as the addition of the calcium channel blocker, lanthanum (10 muM), abolished the 5-HT- and the 5-HT3 receptor agonists'-stimulated phosphoinositide (PI) response. By contrast, the calcium ionophores, A 23187 and Ionomycin (both at 30 muM) stimulated PI hydrolysis, and this effect was additive to the increased PI turnover induced by 5-HT, 2-Me-5-HT and PBG. The increase in phosphoinositide hydrolysis induced by 5-HT and 2-Me-5-HT was significantly inhibited by phorbol dibutyrate (PDBu) and phorbol myristate acetate, indicating that the activation of protein kinase C (PKC) may provide negative feedback to the PI response induced by 5-HT and 2-Me-5-HT. The effect of PDBu on 2-Me-5-HT-stimulated PI metabolism was reversed by the PKC inhibitors, staurosporine, calphostin C and chelerythrine (all at 10 muM), however, Pertussis toxin (0.5 and 1 mug) had no effect on either 5-HT's or 2-Me-5-HT's increased stimulation of PI hydrolysis, suggesting that this response is not associated to a G(i) GTP binding protein.