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. 1990 Sep;9(9):2671–2677. doi: 10.1002/j.1460-2075.1990.tb07452.x

Heterogeneity of Drosophila nicotinic acetylcholine receptors: SAD, a novel developmentally regulated alpha-subunit.

E Sawruk 1, P Schloss 1, H Betz 1, B Schmitt 1
PMCID: PMC551970  PMID: 1697262

Abstract

Two genes, ard and als, are known to encode subunits of the nicotinic acetylcholine receptor (nAChR) in Drosophila. Here we describe the isolation of cDNA clones encoding a novel member (SAD, or alpha 2) of this receptor protein family. The deduced amino acid sequence displays high homology to the ALS protein and shares structural features with ligand binding nAChR alpha-subunits. Sad transcripts accumulate during major periods of neuronal differentiation and, in embryos, are localized in the central nervous system. Expression of SAD cRNA in Xenopus oocytes generates cation channels that are gated by nicotine. These data indicate heterogeneity of nAChRs in Drosophila.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ballivet M., Nef P., Couturier S., Rungger D., Bader C. R., Bertrand D., Cooper E. Electrophysiology of a chick neuronal nicotinic acetylcholine receptor expressed in Xenopus oocytes after cDNA injection. Neuron. 1988 Nov;1(9):847–852. doi: 10.1016/0896-6273(88)90132-8. [DOI] [PubMed] [Google Scholar]
  2. Bertrand D., Ballivet M., Rungger D. Activation and blocking of neuronal nicotinic acetylcholine receptor reconstituted in Xenopus oocytes. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1993–1997. doi: 10.1073/pnas.87.5.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bossy B., Ballivet M., Spierer P. Conservation of neural nicotinic acetylcholine receptors from Drosophila to vertebrate central nervous systems. EMBO J. 1988 Mar;7(3):611–618. doi: 10.1002/j.1460-2075.1988.tb02854.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boulter J., Connolly J., Deneris E., Goldman D., Heinemann S., Patrick J. Functional expression of two neuronal nicotinic acetylcholine receptors from cDNA clones identifies a gene family. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7763–7767. doi: 10.1073/pnas.84.21.7763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boulter J., Evans K., Goldman D., Martin G., Treco D., Heinemann S., Patrick J. Isolation of a cDNA clone coding for a possible neural nicotinic acetylcholine receptor alpha-subunit. 1986 Jan 30-Feb 5Nature. 319(6052):368–374. doi: 10.1038/319368a0. [DOI] [PubMed] [Google Scholar]
  6. Boulter J., O'Shea-Greenfield A., Duvoisin R. M., Connolly J. G., Wada E., Jensen A., Gardner P. D., Ballivet M., Deneris E. S., McKinnon D. Alpha 3, alpha 5, and beta 4: three members of the rat neuronal nicotinic acetylcholine receptor-related gene family form a gene cluster. J Biol Chem. 1990 Mar 15;265(8):4472–4482. [PubMed] [Google Scholar]
  7. Breer H., Kleene R., Hinz G. Molecular forms and subunit structure of the acetylcholine receptor in the central nervous system of insects. J Neurosci. 1985 Dec;5(12):3386–3392. doi: 10.1523/JNEUROSCI.05-12-03386.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Carpenter M. K., Parker I., Miledi R. Changes in messenger RNAs coding for neurotransmitter receptors and voltage-operated channels in the developing rat cerebral cortex. Dev Biol. 1990 Apr;138(2):313–323. doi: 10.1016/0012-1606(90)90199-s. [DOI] [PubMed] [Google Scholar]
  9. Cathala G., Savouret J. F., Mendez B., West B. L., Karin M., Martial J. A., Baxter J. D. A method for isolation of intact, translationally active ribonucleic acid. DNA. 1983;2(4):329–335. doi: 10.1089/dna.1983.2.329. [DOI] [PubMed] [Google Scholar]
  10. Changeux J. P., Devillers-Thiéry A., Chemouilli P. Acetylcholine receptor: an allosteric protein. Science. 1984 Sep 21;225(4668):1335–1345. doi: 10.1126/science.6382611. [DOI] [PubMed] [Google Scholar]
  11. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  12. Deneris E. S., Connolly J., Boulter J., Wada E., Wada K., Swanson L. W., Patrick J., Heinemann S. Primary structure and expression of beta 2: a novel subunit of neuronal nicotinic acetylcholine receptors. Neuron. 1988 Mar;1(1):45–54. doi: 10.1016/0896-6273(88)90208-5. [DOI] [PubMed] [Google Scholar]
  13. Dudai Y., Amsterdam A. Nicotinic receptors in the brain of Drosophila melanogaster demonstrated by autoradiography with [125I]alpha-bungarotoxin. Brain Res. 1977 Jul 22;130(3):551–555. doi: 10.1016/0006-8993(77)90117-2. [DOI] [PubMed] [Google Scholar]
  14. Duvoisin R. M., Deneris E. S., Patrick J., Heinemann S. The functional diversity of the neuronal nicotinic acetylcholine receptors is increased by a novel subunit: beta 4. Neuron. 1989 Oct;3(4):487–496. doi: 10.1016/0896-6273(89)90207-9. [DOI] [PubMed] [Google Scholar]
  15. Goldman D., Deneris E., Luyten W., Kochhar A., Patrick J., Heinemann S. Members of a nicotinic acetylcholine receptor gene family are expressed in different regions of the mammalian central nervous system. Cell. 1987 Mar 27;48(6):965–973. doi: 10.1016/0092-8674(87)90705-7. [DOI] [PubMed] [Google Scholar]
  16. Gration K. A., Lambert J. J., Ramsey R., Usherwood P. N. Non-random openings and concentration-dependent lifetimes of glutamate-gated channels in muscle membrane. Nature. 1981 Jun 4;291(5814):423–425. doi: 10.1038/291423a0. [DOI] [PubMed] [Google Scholar]
  17. Grenningloh G., Rienitz A., Schmitt B., Methfessel C., Zensen M., Beyreuther K., Gundelfinger E. D., Betz H. The strychnine-binding subunit of the glycine receptor shows homology with nicotinic acetylcholine receptors. Nature. 1987 Jul 16;328(6127):215–220. doi: 10.1038/328215a0. [DOI] [PubMed] [Google Scholar]
  18. Grenningloh G., Schmieden V., Schofield P. R., Seeburg P. H., Siddique T., Mohandas T. K., Becker C. M., Betz H. Alpha subunit variants of the human glycine receptor: primary structures, functional expression and chromosomal localization of the corresponding genes. EMBO J. 1990 Mar;9(3):771–776. doi: 10.1002/j.1460-2075.1990.tb08172.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hanke W., Breer H. Channel properties of an insect neuronal acetylcholine receptor protein reconstituted in planar lipid bilayers. Nature. 1986 May 8;321(6066):171–174. doi: 10.1038/321171a0. [DOI] [PubMed] [Google Scholar]
  20. Hermans-Borgmeyer I., Hoffmeister S., Sawruk E., Betz H., Schmitt B., Gundelfinger E. D. Neuronal acetylcholine receptors in Drosophila: mature and immature transcripts of the ard gene in the developing central nervous system. Neuron. 1989 Feb;2(2):1147–1156. doi: 10.1016/0896-6273(89)90181-5. [DOI] [PubMed] [Google Scholar]
  21. Hermans-Borgmeyer I., Zopf D., Ryseck R. P., Hovemann B., Betz H., Gundelfinger E. D. Primary structure of a developmentally regulated nicotinic acetylcholine receptor protein from Drosophila. EMBO J. 1986 Jul;5(7):1503–1508. doi: 10.1002/j.1460-2075.1986.tb04389.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hollmann M., O'Shea-Greenfield A., Rogers S. W., Heinemann S. Cloning by functional expression of a member of the glutamate receptor family. Nature. 1989 Dec 7;342(6250):643–648. doi: 10.1038/342643a0. [DOI] [PubMed] [Google Scholar]
  23. Kao P. N., Karlin A. Acetylcholine receptor binding site contains a disulfide cross-link between adjacent half-cystinyl residues. J Biol Chem. 1986 Jun 25;261(18):8085–8088. [PubMed] [Google Scholar]
  24. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  25. Levitan E. S., Schofield P. R., Burt D. R., Rhee L. M., Wisden W., Köhler M., Fujita N., Rodriguez H. F., Stephenson A., Darlison M. G. Structural and functional basis for GABAA receptor heterogeneity. Nature. 1988 Sep 1;335(6185):76–79. doi: 10.1038/335076a0. [DOI] [PubMed] [Google Scholar]
  26. Lütcke H. A., Chow K. C., Mickel F. S., Moss K. A., Kern H. F., Scheele G. A. Selection of AUG initiation codons differs in plants and animals. EMBO J. 1987 Jan;6(1):43–48. doi: 10.1002/j.1460-2075.1987.tb04716.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Maniatis T., Hardison R. C., Lacy E., Lauer J., O'Connell C., Quon D., Sim G. K., Efstratiadis A. The isolation of structural genes from libraries of eucaryotic DNA. Cell. 1978 Oct;15(2):687–701. doi: 10.1016/0092-8674(78)90036-3. [DOI] [PubMed] [Google Scholar]
  28. Mishina M., Tobimatsu T., Imoto K., Tanaka K., Fujita Y., Fukuda K., Kurasaki M., Takahashi H., Morimoto Y., Hirose T. Location of functional regions of acetylcholine receptor alpha-subunit by site-directed mutagenesis. 1985 Jan 31-Feb 6Nature. 313(6001):364–369. doi: 10.1038/313364a0. [DOI] [PubMed] [Google Scholar]
  29. Nef P., Oneyser C., Alliod C., Couturier S., Ballivet M. Genes expressed in the brain define three distinct neuronal nicotinic acetylcholine receptors. EMBO J. 1988 Mar;7(3):595–601. doi: 10.1002/j.1460-2075.1988.tb02852.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Neumann D., Barchan D., Safran A., Gershoni J. M., Fuchs S. Mapping of the alpha-bungarotoxin binding site within the alpha subunit of the acetylcholine receptor. Proc Natl Acad Sci U S A. 1986 May;83(9):3008–3011. doi: 10.1073/pnas.83.9.3008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Noda M., Takahashi H., Tanabe T., Toyosato M., Furutani Y., Hirose T., Asai M., Inayama S., Miyata T., Numa S. Primary structure of alpha-subunit precursor of Torpedo californica acetylcholine receptor deduced from cDNA sequence. Nature. 1982 Oct 28;299(5886):793–797. doi: 10.1038/299793a0. [DOI] [PubMed] [Google Scholar]
  32. Rudloff E. Acetylcholine receptors in the central nervous system of Drosophila melanogaster. Exp Cell Res. 1978 Jan;111(1):185–190. doi: 10.1016/0014-4827(78)90248-3. [DOI] [PubMed] [Google Scholar]
  33. Sanes J. R., Prescott D. J., Hildebrand J. G. Cholinergic neurochemical development of normal and deafferented antennal lobes during metamorphosis of the moth, Manduca sexta. Brain Res. 1977 Jan 7;119(2):389–402. doi: 10.1016/0006-8993(77)90318-3. [DOI] [PubMed] [Google Scholar]
  34. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sawruk E., Hermans-Borgmeyer I., Betz H., Gundelfinger E. D. Characterization of an invertebrate nicotinic acetylcholine receptor gene: the ard gene of Drosophila melanogaster. FEBS Lett. 1988 Aug 1;235(1-2):40–46. doi: 10.1016/0014-5793(88)81230-4. [DOI] [PubMed] [Google Scholar]
  36. Schloss P., Hermans-Borgmeyer I., Betz H., Gundelfinger E. D. Neuronal acetylcholine receptors in Drosophila: the ARD protein is a component of a high-affinity alpha-bungarotoxin binding complex. EMBO J. 1988 Sep;7(9):2889–2894. doi: 10.1002/j.1460-2075.1988.tb03146.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schmidt-Nielsen B. K., Gepner J. I., Teng N. N., Hall L. M. Characterization of an alpha-bungarotoxin binding component from Drosophila melanogaster. J Neurochem. 1977 Dec;29(6):1013–1029. doi: 10.1111/j.1471-4159.1977.tb06505.x. [DOI] [PubMed] [Google Scholar]
  38. Schmieden V., Grenningloh G., Schofield P. R., Betz H. Functional expression in Xenopus oocytes of the strychnine binding 48 kd subunit of the glycine receptor. EMBO J. 1989 Mar;8(3):695–700. doi: 10.1002/j.1460-2075.1989.tb03428.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Schofield P. R., Darlison M. G., Fujita N., Burt D. R., Stephenson F. A., Rodriguez H., Rhee L. M., Ramachandran J., Reale V., Glencorse T. A. Sequence and functional expression of the GABA A receptor shows a ligand-gated receptor super-family. Nature. 1987 Jul 16;328(6127):221–227. doi: 10.1038/328221a0. [DOI] [PubMed] [Google Scholar]
  40. Stroud R. M., Finer-Moore J. Acetylcholine receptor structure, function, and evolution. Annu Rev Cell Biol. 1985;1:317–351. doi: 10.1146/annurev.cb.01.110185.001533. [DOI] [PubMed] [Google Scholar]
  41. Tautz D., Pfeifle C. A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback. Chromosoma. 1989 Aug;98(2):81–85. doi: 10.1007/BF00291041. [DOI] [PubMed] [Google Scholar]
  42. Wada K., Ballivet M., Boulter J., Connolly J., Wada E., Deneris E. S., Swanson L. W., Heinemann S., Patrick J. Functional expression of a new pharmacological subtype of brain nicotinic acetylcholine receptor. Science. 1988 Apr 15;240(4850):330–334. doi: 10.1126/science.2832952. [DOI] [PubMed] [Google Scholar]
  43. Wadsworth S. C., Rosenthal L. S., Kammermeyer K. L., Potter M. B., Nelson D. J. Expression of a Drosophila melanogaster acetylcholine receptor-related gene in the central nervous system. Mol Cell Biol. 1988 Feb;8(2):778–785. doi: 10.1128/mcb.8.2.778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. White K., Kankel D. R. Patterns of cell division and cell movement in the formation of the imaginal nervous system in Drosophila melanogaster. Dev Biol. 1978 Aug;65(2):296–321. doi: 10.1016/0012-1606(78)90029-5. [DOI] [PubMed] [Google Scholar]
  45. Whiting P., Lindstrom J. Purification and characterization of a nicotinic acetylcholine receptor from rat brain. Proc Natl Acad Sci U S A. 1987 Jan;84(2):595–599. doi: 10.1073/pnas.84.2.595. [DOI] [PMC free article] [PubMed] [Google Scholar]

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