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Volume 17, Number 17, Issue of September 1, 1997 pp. 6545-6553
Copyright ©1997 Society for Neuroscience

Agonist-Specific Coupling of a Cloned Drosophila melanogaster D1-Like Dopamine Receptor to Multiple Second Messenger Pathways by Synthetic Agonists

Received May 6, 1997; revised June 10, 1997; accepted June 12, 1997.

Vincenzina Reale¹, Frances Hannan¹, Linda M. Hall², and Peter D. Evans¹

¹ The Babraham Institute Laboratory of Molecular Signaling, Department of Zoology, University of Cambridge, Cambridge CB2 3EJ, United Kingdom, and ² Department of Biochemical Pharmacology, State University of New York at Buffalo, Buffalo, New York 14260-1200

The mechanism of coupling of a cloned Drosophila D1-like dopamine receptor, DopR99B, to multiple second messenger systems when expressed in Xenopus oocytes is described. The receptor is coupled directly to the generation of a rapid, transient intracellular Ca²⁺ signal, monitored as changes in inward current mediated by the oocyte endogenous Ca²⁺-activated chloride channel, by a pertussis toxin-insensitive G-protein-coupled pathway. The more prolonged receptor-mediated changes in adenylyl cyclase activity are generated by an independent G-protein-coupled pathway that is pertussis toxin-sensitive but calcium-independent, and G-subunits appear to be involved in the transduction of this response. This is the first evidence for the direct coupling of a cloned D1-like dopamine receptor both to the activation of adenylyl cyclase and to the initiation of an intracellular Ca²⁺ signal. The pharmacological profile of both second messenger effects is identical for a range of naturally occurring catecholamine ligands (dopamine > norepinephrine > epinephrine) and for the blockade of dopamine responses by a range of synthetic antagonists. However, the pharmacological profiles of the two second messenger responses differ for a range of synthetic agonists. Thus, the receptor exhibits agonist-specific coupling to second messenger systems for synthetic agonists. This feature could provide a useful tool in the genetic analysis of the roles of the multiple second messenger pathways activated by this receptor, given the likely involvement of dopamine in the processes of learning and memory in the insect nervous system.

Key words: cloned dopamine receptor; Drosophila melanogaster; Xenopus oocyte expression; calcium; adenylyl cyclase; G-protein coupling

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