A primordial dopamine D1-like adenylyl cyclase-linked receptor from Drosophila melanogaster displaying poor affinity for benzazepines

Kim S. Sugamori, Lidia L. Demchyshyn, Fortunata McConkey, Michael Forte, Hyman B. Niznik

Research output: Contribution to journalArticle

114 Citations (Scopus)

Abstract

We report here the isolation from Drosophila melanogaster of a 2.0 kb cDNA clone encoding a 385 amino acid protein (dDA1) displaying, within putative transmembrane domains, highest amino acid sequence homology (49-53%) to members of the vertebrate dopamine D1-like receptor family. When expressed in either Sf9 or COS-7 cells, dDA1 did not bind the specific D1-like receptor antagonist [3H]SCH-23390 or numerous other dopaminergic, adrenergic or serotoninergic ligands with high affinity. However, like vertebrate dopamine D1-like receptors, dDA1 stimulated the accumulation of cAMP in response to DA (EC50 ∼300 nM) and 6,7-ADTN (EC50∼500 nM). The dopaminergic rank order of potency (DA > NE≫5-HT) and the lack of stimulation by other possible neurotransmitters (octopamine, tyramine, tryptamine) or DA metabolites (e.g. N-acetyl dopamine) found in Drosophila suggests that this receptor functionally belongs to the dopamine D1-like subfamily. Benzazepines, which characteristically bind to vertebrate dopamine D1-like receptors with high affinity, were relatively poor in stimulating (SKF-38393, SKF-82526; EC50 > 10 μM) dDA1-mediated accumulation of cAMP. Of the numerous compounds tested, a few dopaminergic antagonists inhibited DA-stimulated production of cAMP in a concentration-dependent manner, albeit with considerably reduced affinity, and with the rank order of potency: (+)-butaclamol(Kb∼125nM) > SCH-23390(Kb∼230nM) > α-flupenthixol (Kb ∼ 400 nM) > chlorpromazine ≥ spiperone (Kb ∼ 680 nM) ≥ clozapine In situ hybridization revealed that dDA1 receptor mRNA is expressed as a maternal transcript, and at later blastoderm stages is restricted to apical regions of the cortical peripheral cytoplasm. The generation of inter-species D1 receptor chimeras may help to identify those particular sequence-specific motifs or amino acid residues confering high affinity benzazepine receptor interactions.

Original languageEnglish (US)
Pages (from-to)131-138
Number of pages8
JournalFEBS Letters
Volume362
Issue number2
DOIs
StatePublished - Apr 3 1995

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Benzazepines
Dopamine D1 Receptors
Drosophila melanogaster
Adenylyl Cyclases
Vertebrates
Dopamine
Butaclamol
Fenoldopam
Flupenthixol
Octopamine
Blastoderm
2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine
Spiperone
Amino Acid Sequence Homology
Amino Acid Motifs
Tyramine
Dopamine Antagonists
Amino Acids
Clozapine
COS Cells

Keywords

  • cAMP
  • Catecholamine Sequences reported in this paper have been deposited in GenBank with Accession Number U22106
  • G protein-coupled receptor
  • Invertebrate

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology
  • Structural Biology
  • Biochemistry
  • Biophysics

Cite this

A primordial dopamine D1-like adenylyl cyclase-linked receptor from Drosophila melanogaster displaying poor affinity for benzazepines. / Sugamori, Kim S.; Demchyshyn, Lidia L.; McConkey, Fortunata; Forte, Michael; Niznik, Hyman B.

In: FEBS Letters, Vol. 362, No. 2, 03.04.1995, p. 131-138.

Research output: Contribution to journalArticle

Sugamori, Kim S. ; Demchyshyn, Lidia L. ; McConkey, Fortunata ; Forte, Michael ; Niznik, Hyman B. / A primordial dopamine D1-like adenylyl cyclase-linked receptor from Drosophila melanogaster displaying poor affinity for benzazepines. In: FEBS Letters. 1995 ; Vol. 362, No. 2. pp. 131-138.
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