Bradykinin and phorbol ester but not 5-HT2B receptor activation stimulate phospholipase D activity in the rat stomach fundus.

1. Serotonin has been implicated as a mediator involved in migraine headache, an effect that may involve central 5-HT2B receptor activation. 5-HT2B receptor signal transduction in controversial. 2. Rat stomach fundus contraction to serotonin has been used as a model for 5-HT2B receptor activation. Serotonin-induced contractility involves intracellular calcium release and activation of protein kinase C without stimulation of phosphoinositide (PI) hydrolysis. 3. Since phospholipase D (PLD) activation results in phosphatidic acid production, which can release intracellular calcium and provide diacylglycerol for PKC activation, the purpose of this study was to determine whether the 5-HT2B receptor coupled to PLD activation using the rat stomach fundus as a model system. 4. Using phosphatidylethanol production to measure PLD activity, both bradykinin (0.01-1 microM) and phorbol dibutyrate (PDBu, 1 microM) stimulated PLD activity in rat stomach fundal strips, indicating that this tissue possesses an active PLD system. 5. Under identical conditions, 5-hydroxytryptamine (5-HT) failed to stimulate PLD activity over a concentration range (1 nM-1 microM) documented to induce 5-HT2B receptor-mediated contraction in rat stomach fundus. Thus, the 5-HT2B contractile receptor in rat stomach fundus is not coupled to PLD activation, whereas both bradykinin and phorbol ester do couple to PLD.