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Characterisation of AmphiAmR4, an amphioxus (Branchiostoma floridae) α2-adrenergic-like G-protein-coupled receptor

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Invertebrate Neuroscience

Abstract

Little is known about the evolutionary relationship between vertebrate adrenergic receptors and invertebrate octopamine and tyramine receptors. The complexity of the adrenergic signalling system is believed to be an innovation of the vertebrate lineage but the presence of noradrenaline has been reported in some invertebrate species. The cephalochordate, amphioxus (Branchiostoma floridae), is an ideal model organism for studying the evolution of vertebrate GPCRs, given its unique position at the base of the chordate lineage. Here, we describe the pharmacological characterisation and second messenger coupling abilities of AmphiAmR4, which clusters with α2-adrenergic receptors in a phylogenetic tree but also shares a high sequence similarity to invertebrate octopamine/tyramine receptors in both BLAST and Hidden Markov Model analyses. Thus, it was of particular interest to determine if AmphiAmR4 displayed similar functional properties to the vertebrate α2-adrenergic receptors or to invertebrate octopamine or tyramine receptors. When stably expressed in Chinese hamster ovary (CHO) cells, noradrenaline couples the receptor to both the activation of adenylyl cyclase and to the activation of the MAPKinase pathway. Pharmacological studies with a wide range of agonists and antagonists suggest that AmphiAmR4 functions as an α2-adrenergic-like receptor when expressed in CHO cells.

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Abbreviations

CHO:

Chinese hamster ovary

D-PBS:

Dulbecco’s phosphate-buffered saline

ERK:

Extracellular signal-related kinase

FBS:

Foetal bovine serum

GFP:

Green fluorescent protein

GPCR:

G-protein-coupled receptor

5-HT:

5-Hydroxytryptamine

IBMX:

3-Isobutyl methyl xanthine

MAPK:

Mitogen-activated protein kinase

ORF:

Open reading frame

TM:

Transmembrane domain

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Acknowledgments

This work was supported by the BBSRC through the Babraham Institute and by a BBSRC Studentship to AB. We thank Dr. Mikhail Matz, Whitney Laboratory, University of Florida, St. Augustine, USA for kindly supplying us with the amphioxus cDNA libraries used in this study.

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The authors declare no conflicts of interest.

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  1. The Inositide Laboratory, The Babraham Institute, The Babraham Research Campus, Cambridge, CB22 3AT, UK

    Asha Bayliss & Peter D. Evans

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  1. Asha Bayliss
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  2. Peter D. Evans
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Correspondence to Peter D. Evans.

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Bayliss, A., Evans, P.D. Characterisation of AmphiAmR4, an amphioxus (Branchiostoma floridae) α2-adrenergic-like G-protein-coupled receptor. Invert Neurosci 13, 71–84 (2013). https://doi.org/10.1007/s10158-012-0145-6

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