Abstract
Muscarinic acetylcholine receptors (mAChRs) play a central role in the mammalian nervous system. These receptors are G protein-coupled receptors (GPCRs), which are activated by the agonists acetylcholine and muscarine, and blocked by a variety of antagonists. Mammals have five mAChRs (m1–m5). In this study, we cloned two structurally related GPCRs from the fruit fly Drosophila melanogaster, which, after expression in Chinese hamster ovary cells, proved to be muscarinic acetylcholine receptors. One mAChR (the A-type; encoded by gene CG4356) is activated by acetylcholine (EC50, 5 × 10−8 M) and muscarine (EC50, 6 × 10−8 M) and blocked by the classical mAChR antagonists atropine, scopolamine, and 3-quinuclidinyl-benzilate (QNB), while the other (the B-type; encoded by gene CG7918) is also activated by acetylcholine, but has a 1,000-fold lower sensitivity to muscarine, and is not blocked by the antagonists. A- and B-type mAChRs were also cloned and functionally characterized from the red flour beetle Tribolium castaneum. Recently, Haga et al. (Nature 2012, 482: 547–551) published the crystal structure of the human m2 mAChR, revealing 14 amino acid residues forming the binding pocket for QNB. These residues are identical between the human m2 and the D. melanogaster and T. castaneum A-type mAChRs, while many of them are different between the human m2 and the B-type receptors. Using bioinformatics, one orthologue of the A-type and one of the B-type mAChRs could also be found in all other arthropods with a sequenced genome. Protostomes, such as arthropods, and deuterostomes, such as mammals and other vertebrates, belong to two evolutionarily distinct lineages of animal evolution that split about 700 million years ago. We found that animals that originated before this split, such as cnidarians (Hydra), had two A-type mAChRs. From these data we propose a model for the evolution of mAChRs.
Similar content being viewed by others
Abbreviations
- CHO:
-
Chinese hamster ovary
- mAChR:
-
Muscarinic acetylcholine receptor
- MYR:
-
Million years
- QNB:
-
3-Quinuclidinyl-benzylate
- qPCR:
-
Quantitative PCR
References
Douzéry EJ, Snell EA, Bapteste E, Delsuc F, Philippe H (2004) The timing of eukaryotic evolution: does a relaxed molecular clock reconcile proteins and fossils? Proc Natl Acad Sci USA 101:15386–15391
Yakel JL (2010) Gating of nicotinic Ach receptors: latest insights into ligand binding and function. J Physiol 588:597–602
Millar NS, Harkness PC (2008) Assembly and trafficking of nicotinic acetylcholine receptors (Review). Mol Membr Biol 25:279–292
Jones AK, Sattelle DB (2010) Diversity of insect nicotinic acetylcholine receptor subunits. Adv Exp Med Biol 683:25–43
Dupuis J, Louis T, Gauthier M, Raymond V (2012) Insights from honeybee (Apis mellifera) and fly (Drosophila melanogaster) nicotinic acetylcholine receptors: from genes to behavioral functions. Neurosci Biobehav Rev 36:1553–1564
Bubser M, Byun N, Wood MR, Jones CK (2012) Muscarinic receptor pharmacology and circuitry for the modulation of cognition. Hanb Exp Pharmacol 208:121–166
Tobin G, Giglio D, Lundgren O (2009) Muscarinic receptor subtypes in the alimentary tract. J Physiol Pharmacol 60:3–21
Harvey RD (2012) Muscarinic receptor agonists and antagonists: effects on cardiovascular function. Hanb Exp Pharmacol 208:299–316
Shapiro RA, Wakimoto BT, Subers EM, Nathanson NM (1989) Characterization and functional expression in mammalian cells of genomic and cDNA clones encoding a Drosophila muscarinic acetylcholine receptor. Proc Natl Acad Sci USA 86:9039–9043
Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-CPR data by geometric averaging of multiple internal control genes. Genome Biol 3:R0034.1–R0034.11
Stables J, Green A, Marshall F, Fraser N, Knight E, Sautel M, Milligan G, Lee M, Rees S (1997) A bioluminescent assay for agonist activity at potentially any G-protein-coupled receptor. Anal Biochem 252:115–126
Secher T, Lenz C, Cazzamali G, Sørensen G, Williamson M, Hansen GN, Svane P, Grimmelikhuijzen CJP (2001) Molecular cloning of a functional allatostatin gut/brain receptor and an allatostatin preprohormone from the silkworm Bombyx mori. J Biol Chem 276:47052–47060
Staubli F, Jørgensen TJD, Cazzamali G, Williamson M, Lenz C, Søndergaard L, Roepstorff P, Grimmelikhuijzen CJP (2002) Molecular identification of the insect adipokinetic hormone receptors. Proc Natl Acad Sci USA 99:3446–3451
Millar NS, Baylis HA, Reaper C, Bunting R, Mason WT, Sattelle DB (1995) Functional expression of a cloned Drosophila muscarinic acetylcholine receptor in a stable Drosophila cell line. J Exp Biol 198:1843–1850
Onai T, FitzGerald MG, Arakawa S, Gocayne JD, Urquhart DA, Hall LM, Fraser CM, McCombie WR, Venter JC (1989) Cloning, sequence analysis and chromosome localization of a Drosophila muscarinic acetylcholine receptor. FEBS Lett 255:219–225
Brody T, Cravchik A (2000) Drosophila melanogaster G protein-coupled receptors. J Cell Biol 150:F83–F88
Hauser F, Cazzamali G, Williamson M, Blenau W, Grimmelikhuijzen CJP (2006) A review of neurohormone GPCRs present in the fruitfly Drosophila melanogaster and the honey bee Apis mellifera. Prog Neurobiol 80:1–19
Hauser F, Cazzamali G, Williamson M, Park Y, Li B, Tanaka Y, Predel R, Neupert S, Schachtner J, Verleyen P, Grimmelikhuijzen CJP (2008) A genome-wide inventory of neurohormone GPCRs in the red flour beetle Tribolium castaneum. Front Neuroendocrinol 29:142–165
Haga K, Kruse AC, Asada H, Yurugi-Kobayashi T, Shiroishi M, Zhang C, Weis WI, Okada T, Kobilka BK, Haga T, Kobayashi T (2012) Structure and function of the human M2 muscarinic acetylcholine receptor bound to an antagonist. Nature 482:547–551
Kruse AC, Hu J, Pan AC, Arlow DH, Rosenbaum DM, Rosemond E, Green HF, Liu T, Chae PS, Dror RO, Shaw DE, Weis WI, Wess J, Kobilka BK (2012) Structure and dynamics of the M3 muscarinic acetylcholine receptor. Nature 482:552–556
Blüml K, Mutschler E, Wess J (1994) Functional role in ligand binding and receptor activation of an asparagine residue present in the sixth transmembrane domain of all muscarinic acetylcholine receptors. J Biol Chem 269:18870–18876
Ward SD, Curtis CA, Hulme EC (1999) Alanine-scanning mutagenesis of transmembrane domain 6 of the M(1) muscarinic acetylcholine receptor suggests that Tyr381 plays key roles in receptor function. Mol Pharmacol 56:1031–1041
Lee Y-S, Park Y-S, Chang DJ, Hwang JM, Min CK, Kaang B-K, Cho NJ (1999) Cloning and expression of a G protein-linked acetylcholine receptor from Caenorhabditis elegans. J Neurochem 72:58–65
Lee Y-S, Park Y-S, Nam S, Suh S-J, Lee J, Kaang B-K, Cho NJ (2000) Characterization of GAR-2, a novel G protein-linked acetylcholine receptor from Caenorhabditis elegans. J Neurochem 75:1800–1809
Park Y-S, Cho T-J, Cho NJ (2006) Stimulation of cyclic AMP production by the Caenorhabditis elegans muscarinic acetylcholine receptor GAR-3 in Chinese hamster ovary cells. Arch Biochem Biophys 450:203–207
Acknowledgments
We thank Johannes Thomsen for ty** the manuscript, and the Danish Research Agency, Novo Nordisk Foundation, and Carlsberg Foundation for financial support.
Author information
Authors and Affiliations
Corresponding author
Additional information
Data deposition: The sequences reported in this paper have been deposited in the GenBank database (accession nos. JQ860106, JQ860107, JQ922420, JQ922421, JX028234, JX028235, JX174094).
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Collin, C., Hauser, F., de Valdivia, E.G. et al. Two types of muscarinic acetylcholine receptors in Drosophila and other arthropods. Cell. Mol. Life Sci. 70, 3231–3242 (2013). https://doi.org/10.1007/s00018-013-1334-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00018-013-1334-0