Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Feb 1;91(3):1188–1192. doi: 10.1073/pnas.91.3.1188

A cocaine-sensitive Drosophila serotonin transporter: cloning, expression, and electrophysiological characterization.

J L Corey 1, M W Quick 1, N Davidson 1, H A Lester 1, J Guastella 1
PMCID: PMC521479  PMID: 8302852

Abstract

A cocaine-sensitive, high-affinity Drosophila serotonin (5-hydroxytryptamine; 5HT) transporter cDNA, denoted dSERT1, was isolated and characterized in oocytes. dSERT1 shows little transport of other monoamines and is Na+ and Cl- dependent. Sequence analysis indicates 12 putative transmembrane domains and strong homologies (approximately 50%) among dSERT1 and mammalian 5HT, norepinephrine, and dopamine transporters. Interestingly, the pharmacological properties of dSERT1, including sensitivity to antidepressants, are more similar to those of mammalian catecholamine transporters than to mammalian 5HT transporters. Two-electrode voltage-clamp analysis demonstrated 5HT-induced, voltage-dependent currents. Cloning and characterization of dSERT1 adds significantly to our knowledge of the diversity of 5HT transporters with regard to primary sequence, pharmacological profile, and permeation properties.

Full text

PDF
1188

Selected References

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

  1. Amara S. G., Kuhar M. J. Neurotransmitter transporters: recent progress. Annu Rev Neurosci. 1993;16:73–93. doi: 10.1146/annurev.ne.16.030193.000445. [DOI] [PubMed] [Google Scholar]
  2. Arriza J. L., Kavanaugh M. P., Fairman W. A., Wu Y. N., Murdoch G. H., North R. A., Amara S. G. Cloning and expression of a human neutral amino acid transporter with structural similarity to the glutamate transporter gene family. J Biol Chem. 1993 Jul 25;268(21):15329–15332. [PubMed] [Google Scholar]
  3. Blakely R. D., Berson H. E., Fremeau R. T., Jr, Caron M. G., Peek M. M., Prince H. K., Bradley C. C. Cloning and expression of a functional serotonin transporter from rat brain. Nature. 1991 Nov 7;354(6348):66–70. doi: 10.1038/354066a0. [DOI] [PubMed] [Google Scholar]
  4. Borden L. A., Smith K. E., Hartig P. R., Branchek T. A., Weinshank R. L. Molecular heterogeneity of the gamma-aminobutyric acid (GABA) transport system. Cloning of two novel high affinity GABA transporters from rat brain. J Biol Chem. 1992 Oct 15;267(29):21098–21104. [PubMed] [Google Scholar]
  5. Borowsky B., Mezey E., Hoffman B. J. Two glycine transporter variants with distinct localization in the CNS and peripheral tissues are encoded by a common gene. Neuron. 1993 May;10(5):851–863. doi: 10.1016/0896-6273(93)90201-2. [DOI] [PubMed] [Google Scholar]
  6. Buchner E., Bader R., Buchner S., Cox J., Emson P. C., Flory E., Heizmann C. W., Hemm S., Hofbauer A., Oertel W. H. Cell-specific immuno-probes for the brain of normal and mutant Drosophila melanogaster. I. Wildtype visual system. Cell Tissue Res. 1988 Aug;253(2):357–370. doi: 10.1007/BF00222292. [DOI] [PubMed] [Google Scholar]
  7. Buchner E. Genes expressed in the adult brain of Drosophila and effects of their mutations on behavior: a survey of transmitter- and second messenger-related genes. J Neurogenet. 1991;7(4):153–192. doi: 10.3109/01677069109167432. [DOI] [PubMed] [Google Scholar]
  8. Clark J. A., Deutch A. Y., Gallipoli P. Z., Amara S. G. Functional expression and CNS distribution of a beta-alanine-sensitive neuronal GABA transporter. Neuron. 1992 Aug;9(2):337–348. doi: 10.1016/0896-6273(92)90172-a. [DOI] [PubMed] [Google Scholar]
  9. Fremeau R. T., Jr, Caron M. G., Blakely R. D. Molecular cloning and expression of a high affinity L-proline transporter expressed in putative glutamatergic pathways of rat brain. Neuron. 1992 May;8(5):915–926. doi: 10.1016/0896-6273(92)90206-s. [DOI] [PubMed] [Google Scholar]
  10. Giros B., el Mestikawy S., Bertrand L., Caron M. G. Cloning and functional characterization of a cocaine-sensitive dopamine transporter. FEBS Lett. 1991 Dec 16;295(1-3):149–154. doi: 10.1016/0014-5793(91)81406-x. [DOI] [PubMed] [Google Scholar]
  11. Guastella J., Brecha N., Weigmann C., Lester H. A., Davidson N. Cloning, expression, and localization of a rat brain high-affinity glycine transporter. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7189–7193. doi: 10.1073/pnas.89.15.7189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guastella J., Nelson N., Nelson H., Czyzyk L., Keynan S., Miedel M. C., Davidson N., Lester H. A., Kanner B. I. Cloning and expression of a rat brain GABA transporter. Science. 1990 Sep 14;249(4974):1303–1306. doi: 10.1126/science.1975955. [DOI] [PubMed] [Google Scholar]
  13. Guastella J., Stretton A. O. Distribution of 3H-GABA uptake sites in the nematode Ascaris. J Comp Neurol. 1991 May 22;307(4):598–608. doi: 10.1002/cne.903070407. [DOI] [PubMed] [Google Scholar]
  14. Guimbal C., Kilimann M. W. A Na(+)-dependent creatine transporter in rabbit brain, muscle, heart, and kidney. cDNA cloning and functional expression. J Biol Chem. 1993 Apr 25;268(12):8418–8421. [PubMed] [Google Scholar]
  15. Hamilton B. A., Palazzolo M. J., Meyerowitz E. M. Rapid isolation of long cDNA clones from existing libraries. Nucleic Acids Res. 1991 Apr 25;19(8):1951–1952. doi: 10.1093/nar/19.8.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hoffman B. J., Mezey E., Brownstein M. J. Cloning of a serotonin transporter affected by antidepressants. Science. 1991 Oct 25;254(5031):579–580. doi: 10.1126/science.1948036. [DOI] [PubMed] [Google Scholar]
  17. Kain K. C., Orlandi P. A., Lanar D. E. Universal promoter for gene expression without cloning: expression-PCR. Biotechniques. 1991 Mar;10(3):366–374. [PubMed] [Google Scholar]
  18. Kanai Y., Hediger M. A. Primary structure and functional characterization of a high-affinity glutamate transporter. Nature. 1992 Dec 3;360(6403):467–471. doi: 10.1038/360467a0. [DOI] [PubMed] [Google Scholar]
  19. Kanner B. I., Bendahan A. Transport of 5-hydroxytryptamine in membrane vesicles from rat basophilic leukemia cells. Biochim Biophys Acta. 1985 Jun 27;816(2):403–410. doi: 10.1016/0005-2736(85)90508-5. [DOI] [PubMed] [Google Scholar]
  20. Kilty J. E., Lorang D., Amara S. G. Cloning and expression of a cocaine-sensitive rat dopamine transporter. Science. 1991 Oct 25;254(5031):578–579. doi: 10.1126/science.1948035. [DOI] [PubMed] [Google Scholar]
  21. Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987 Oct 26;15(20):8125–8148. doi: 10.1093/nar/15.20.8125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  23. Liu Q. R., Nelson H., Mandiyan S., López-Corcuera B., Nelson N. Cloning and expression of a glycine transporter from mouse brain. FEBS Lett. 1992 Jun 29;305(2):110–114. doi: 10.1016/0014-5793(92)80875-h. [DOI] [PubMed] [Google Scholar]
  24. Mager S., Naeve J., Quick M., Labarca C., Davidson N., Lester H. A. Steady states, charge movements, and rates for a cloned GABA transporter expressed in Xenopus oocytes. Neuron. 1993 Feb;10(2):177–188. doi: 10.1016/0896-6273(93)90309-f. [DOI] [PubMed] [Google Scholar]
  25. Orkand P. M., Kravitz E. A. Localization of the sites of gamma-aminobutyric acid (GABA) uptake in lobster nerve-muscle preparations. J Cell Biol. 1971 Apr;49(1):75–89. doi: 10.1083/jcb.49.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pacholczyk T., Blakely R. D., Amara S. G. Expression cloning of a cocaine- and antidepressant-sensitive human noradrenaline transporter. Nature. 1991 Mar 28;350(6316):350–354. doi: 10.1038/350350a0. [DOI] [PubMed] [Google Scholar]
  27. Pines G., Danbolt N. C., Bjørås M., Zhang Y., Bendahan A., Eide L., Koepsell H., Storm-Mathisen J., Seeberg E., Kanner B. I. Cloning and expression of a rat brain L-glutamate transporter. Nature. 1992 Dec 3;360(6403):464–467. doi: 10.1038/360464a0. [DOI] [PubMed] [Google Scholar]
  28. Pletscher A. The 5-hydroxytryptamine system of blood platelets: physiology and pathophysiology. Int J Cardiol. 1987 Feb;14(2):177–188. doi: 10.1016/0167-5273(87)90007-6. [DOI] [PubMed] [Google Scholar]
  29. Ramamoorthy S., Bauman A. L., Moore K. R., Han H., Yang-Feng T., Chang A. S., Ganapathy V., Blakely R. D. Antidepressant- and cocaine-sensitive human serotonin transporter: molecular cloning, expression, and chromosomal localization. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2542–2546. doi: 10.1073/pnas.90.6.2542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rudnick G. Active transport of 5-hydroxytryptamine by plasma membrane vesicles isolated from human blood platelets. J Biol Chem. 1977 Apr 10;252(7):2170–2174. [PubMed] [Google Scholar]
  31. Segonzac A., Tateishi T., Langer S. Z. Saturable uptake of [3H]-tryptamine in rabbit platelets is inhibited by 5-hydroxytryptamine uptake blockers. Naunyn Schmiedebergs Arch Pharmacol. 1984 Nov;328(1):33–37. doi: 10.1007/BF00496102. [DOI] [PubMed] [Google Scholar]
  32. Shafqat S., Tamarappoo B. K., Kilberg M. S., Puranam R. S., McNamara J. O., Guadaño-Ferraz A., Fremeau R. T., Jr Cloning and expression of a novel Na(+)-dependent neutral amino acid transporter structurally related to mammalian Na+/glutamate cotransporters. J Biol Chem. 1993 Jul 25;268(21):15351–15355. [PubMed] [Google Scholar]
  33. Shimada S., Kitayama S., Lin C. L., Patel A., Nanthakumar E., Gregor P., Kuhar M., Uhl G. Cloning and expression of a cocaine-sensitive dopamine transporter complementary DNA. Science. 1991 Oct 25;254(5031):576–578. doi: 10.1126/science.1948034. [DOI] [PubMed] [Google Scholar]
  34. Smith K. E., Borden L. A., Hartig P. R., Branchek T., Weinshank R. L. Cloning and expression of a glycine transporter reveal colocalization with NMDA receptors. Neuron. 1992 May;8(5):927–935. doi: 10.1016/0896-6273(92)90207-t. [DOI] [PubMed] [Google Scholar]
  35. Smith K. E., Borden L. A., Wang C. H., Hartig P. R., Branchek T. A., Weinshank R. L. Cloning and expression of a high affinity taurine transporter from rat brain. Mol Pharmacol. 1992 Oct;42(4):563–569. [PubMed] [Google Scholar]
  36. Storck T., Schulte S., Hofmann K., Stoffel W. Structure, expression, and functional analysis of a Na(+)-dependent glutamate/aspartate transporter from rat brain. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10955–10959. doi: 10.1073/pnas.89.22.10955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Usdin T. B., Mezey E., Chen C., Brownstein M. J., Hoffman B. J. Cloning of the cocaine-sensitive bovine dopamine transporter. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11168–11171. doi: 10.1073/pnas.88.24.11168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Vallés A. M., White K. Development of serotonin-containing neurons in Drosophila mutants unable to synthesize serotonin. J Neurosci. 1986 May;6(5):1482–1491. doi: 10.1523/JNEUROSCI.06-05-01482.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wierenga J. M., Hollingworth R. M. Octopamine uptake and metabolism in the insect nervous system. J Neurochem. 1990 Feb;54(2):479–489. doi: 10.1111/j.1471-4159.1990.tb01897.x. [DOI] [PubMed] [Google Scholar]
  40. Wood M. D., Broadhurst A. M., Wyllie M. G. Examination of the relationship between the uptake system for 5-hydroxytryptamine and the high-affinity [3H]imipramine binding site--I. Inhibition by drugs. Neuropharmacology. 1986 May;25(5):519–525. doi: 10.1016/0028-3908(86)90178-4. [DOI] [PubMed] [Google Scholar]
  41. Yamauchi A., Uchida S., Kwon H. M., Preston A. S., Robey R. B., Garcia-Perez A., Burg M. B., Handler J. S. Cloning of a Na(+)- and Cl(-)-dependent betaine transporter that is regulated by hypertonicity. J Biol Chem. 1992 Jan 5;267(1):649–652. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES