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Volume 16, Number 12, Issue of June 15, 1996 pp. 3925-3933
Copyright ©1996 Society for Neuroscience

Cloning and Functional Characterization of a Novel Dopamine Receptor from Drosophila melanogaster

Received Nov. 30, 1995; revised March 26, 1996; accepted April 2, 1996.

Guoping Feng¹, Frances Hannan^{1, 2}, Vincenzina Reale², Yuen Yi Hon¹, Christopher T. Kousky¹, Peter D. Evans², and Linda M. Hall¹

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

A cDNA clone is described that encodes a novel G-protein-coupled dopamine receptor (DopR99B) expressed in Drosophila heads. The DopR99B receptor maps to 99B3-5, close to the position of the octopamine/tyramine receptor gene at 99A10-B1, suggesting that the two may be related through a gene duplication. Agonist stimulation of DopR99B receptors expressed in Xenopus oocytes increased intracellular Ca²⁺ levels monitored as changes in an endogenous inward Ca²⁺-dependent chloride current. In addition to initiating this intracellular Ca²⁺ signal, stimulation of DopR99B increased cAMP levels. The rank order of potency of agonists in stimulating the chloride current is: dopamine > norepinephrine > epinephrine > tyramine. Octopamine and 5-hydroxytryptamine are not active (<100 µM). This pharmacological profile plus the second-messenger coupling pattern suggest that the DopR99B receptor is a D1-like dopamine receptor. However, the hydrophobic core region of the DopR99B receptor shows almost equal amino acid sequence identity (40-48%) with vertebrate serotonergic, 1- and -adrenergic, and D1-like and D2-like dopaminergic receptors. Thus, this Drosophila receptor defines a novel structural class of dopamine receptors. Because DopR99B is the second dopamine receptor cloned from Drosophila, this work establishes dopamine receptor diversity in a system amenable to genetic dissection.

Key words: cloned dopamine receptor; Drosophila melanogaster; Xenopus oocyte expression; adenylyl cyclase; calcium; gene mapping

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