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The Journal of Neuroscience, August 1, 1998, 18(15):5586-5593

Cloning and Functional Expression of an Aplysia 5-HT Receptor Negatively Coupled to Adenylate Cyclase

Annie Angers¹, Maksim V. Storozhuk², Thomas Duchaîne¹, Vincent F. Castellucci^{2, 3}, and Luc DesGroseillers^{1, 3}

¹ Département de biochimie, ² Département de physiologie, and ³ Centre de recherches en sciences neurologiques, Université de Montréal, Montréal, Québec, Canada H3C 3J7

Serotonin (5-HT) is involved in the control of various behaviors in Aplysia californica, including reproduction, feeding, locomotion, circadian rhythm, synaptic plasticity, and synaptic growth. The large variety of functions of 5-HT is mediated by different receptor subtypes that are coupled to different second-messenger systems. Here, we report the cloning of a cDNA coding for an Aplysia G-protein-coupled 5-HT receptor (5-HT_ap1). Its deduced amino acid sequence resembles those of the 5-HT₁ receptor subfamily. When expressed in stable cell lines, 5-HT_ap1 exhibits high-affinity binding for the serotonergic radioligand [N-methyl-³H]lysergic acid diethylamide. This binding is competed by several 5-HT agonists and antagonists, and the pharmacological profile of inhibition has some similarities with those of 5-HT₁ and 5-HT₇ receptors. Application of 5-HT or its agonists 5-carboxamidotryptamine maleate and (±)-8-hydroxy-2-(di-n-propyl-amino) tetralin hydrobromide on cells transformed with 5-HT_ap1 produced a dose-dependent inhibition of forskolin-stimulated cAMP accumulation. 5-HT_ap1 is thus negatively coupled to adenylate cyclase. The production of antiserum against the 5-HT_ap1 receptor allowed us to examine its expression in animal tissues. The receptor protein is detected in every tissue examined, although it seems only weakly expressed in some samples. The receptor is also found in every ganglia of the nervous system, both in the sheath and in the neurons. 5-HT_ap1 mRNA is absent from the sheath, indicating that the protein observed there is probably located on the nerve terminals.

Key words: Aplysia; 5-HT; 5-HT₁ receptor; G-protein-coupled receptor; adenylate cyclase; inhibition

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Copyright © 1998 Society for Neuroscience  0270-6474/98/18155586-08$05.00/0

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