Pharmacological and molecular evidence that the contractile response to serotonin in rat stomach fundus is not mediated by activation of the 5-hydroxytryptamine1C receptor.
The receptor mediating contraction in response to serotonin in the rat stomach fundus has not been characterized in light of the currently acceptable serotonergic receptor classification scheme. Several biochemical and pharmacological approaches to a characterization of this receptor have demonstrated nonidentity with the 5-hydroxytryptamine2 (5HT2), 5HT3, 5HT1A, and 5HT1B receptors, as defined by radiolabeled ligand binding studies in brain cortical membranes. Although there have been reports suggesting that the receptor in the rat stomach fundus may be analogous to the 5HT1C receptor, other pharmacological and biochemical studies have not been consistent with this idea. The present study utilized high affinity ligands for the 5HT1C receptor and the recently derived 5HT1C receptor cDNA clone to provide a more definitive approach to the examination of the relationship between the 5HT1C receptor and the serotonergic contractile receptor in the rat stomach fundus. Using three ligands with high affinity at the 5HT1C receptor, LY53857, ritanserin, and SCH23390, the contractile response to serotonin was inhibited by all three ligands. However, inhibition did not appear competitive nor was the inhibitory potency of these ligands consistent with their affinity at 5HT1C binding sites in brain cortical membranes. We further showed that SCH23390, unlike LY53857 and ritanserin, was also a partial agonist, producing a maximal contraction that was approximately 50% of the maximal response to serotonin in the rat stomach fundus. Thus, the use of these ligands did not support the contention that the receptor mediating serotonin-induced contractions in the rat stomach is identical to the 5HT1C receptor. In more definitive studies using a 5HT1C receptor cDNA probe, we were unable to detect hybridization of the probe with any mRNA species from the rat stomach fundus, whereas the 5HT1C receptor cDNA probe did hybridize to the 5HT1C receptor mRNA in rat brain. Because the cathepsin-D cDNA probe hybridized equally in rat brain and stomach fundus, ensuring the integrity of the RNA preparation from both tissues, the absence of measurable quantities of the 5HT1C receptor mRNA in the rat stomach was probe specific and not an artifact. Furthermore, primers specific for the rat 5HT1C receptor sequence did not detect significant levels of receptor mRNA in rat fundus, although the target sequence was amplified a minimum of 10(5)-fold in a polymerase chain reaction. These studies do not support the contention that the receptor mediating contractile responses to serotonin in the rat stomach fundus is identical to the 5HT1C receptor.(ABSTRACT TRUNCATED AT 400 WORDS)
Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285.
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Cohen
D000818:Animals; D001552:Benzazepines / Q000494:pharmacology; D002402:Cathepsin D / Q000235:genetics; D015342:DNA Probes; D005748:Gastric Fundus / Q000187:drug effects* / Q000502:physiology; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D009119:Muscle Contraction / Q000187:drug effects*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D011985:Receptors, Serotonin / Q000187:drug effects* / Q000235:genetics; D012701:Serotonin / Q000494:pharmacology*; D012702:Serotonin Antagonists / Q000494:pharmacology
D001552:Benzazepines; D015342:DNA Probes; D012333:RNA, Messenger; D011985:Receptors, Serotonin; D012702:Serotonin Antagonists; D012701:Serotonin; D002402:Cathepsin D