SEROTONIN-INDUCED INCREASE IN CAMP IN GANGLIA ISOLATED FROM THE MYENTERIC PLEXUS OF THE GUINEA-PIG SMALL-INTESTINE - MEDIATION BY A NOVEL 5-HT RECEPTOR

Serotonin (5-HT) is a mediator (through 5-HT1P receptors) of slow EPSPs in myenteric ganglia of the small intestine. The effect of 5-HT can be mimicked by elevating cAMP; therefore, we tested the hypothesis that the slow EPSP-like response to 5-HT is cAMP-mediated. Guinea pig gut was enzymatically dissociated; myenteric ganglia remained intact and were collected by filtration. Neurons in the isolated ganglia retained their ability to manifest the slow EPSP-like response to 5-HT. Exposure to 5-HT raised the ganglionic level of cAMP (ED50 0.3 muM). This effect was not antagonized by the 5-HT1P antagonist, N-acetyl-5-hydroxytryptophyl-5-hydroxytryptophan amide (100.0 muM), or mimicked by the 5-HT1P agonist, 5-hydroxyindalpine (10.0 muM). Increases in cAMP were also evoked by the 5-HT1P agonist, 5-carboxyamidotryptamine (10.0 muM), the 5-HT2 agonist, (+/-)1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI; 1.0-10.0 muM), and by the 5-HT4 agonists, renzapride (1.0-10.0 muM) and 5-methoxytryptamine (1.0-10.0 muM); however, neither the 5-HT1/5-HT2 antagonists, spiperone, methysergide, and methiothepin, nor the 5-HT4 antagonist, tropisetron (ICS 205-930; 10.0 muM), were able to inhibit the rise in cAMP evoked by these compounds or by 5-HT (0.1-10.0 muM). The 5-HT-evoked elevation of cAMP was antagonized by ketanserin (10.0 muM), which also blocked the effects of 5-methoxytryptamine and DOI, but not those of renzapride. The effective concentration of DOI, however, was higher than that needed for activation of 5-HT2 receptors, and Northern analysis using a cDNA probe encoding the rat 5-HT2 receptor failed to reveal the presence of 5-HT2 mRNA in myenteric ganglia, although it hybridizes with mRNA of the right size in the guinea pig brain. Compounds that failed to change levels of cAMP or to antagonize the action of 5-HT included 8-hydroxy-di-n-propylamino tetralin, R58639, R88226, and sumatriptan. It is concluded that the receptor responsible for the 5-HT-induced rise in cAMP in ganglia isolated from the guinea pig myenteric plexus is not a known subtype of 5-HT receptor. Since the pharmacology of this novel receptor is different from that of the slow EPSP-like response to 5-HT, the receptor probably does not mediate the slow EPSP.