Characterization of a genetically reconstituted high-affinity system for serotonin transport.
- 1 December 1989
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 86 (23), 9611-9615
- https://doi.org/10.1073/pnas.86.23.9611
Abstract
By transfecting mouse fibroblast L-M cells with human genomic DNA, we have established and identified several clonal cell lines that stably express a high-affinity serotonin (5-HT)-uptake mechanism absent in untransfected host cells. One such cell line, L-S1, possesses features of 5-[3H]HT uptake similar to those previously characterized in the central nervous system and blood platelets: (i) specificity for 5-HT; (ii) antagonism by imipramine, a known inhibitor of high-affinity 5-HT uptake; (iii) both Na+ and temperature dependences; (iv) kinetic saturability; and (v) high affinity for 5-HT (Km = 0.39 +/- 0.10 microM; Vmax = 2.14 +/- 0.55 pmol/min per mg of protein). This cell line can be used to compare the relative efficacies of known blockers of 5-HT uptake and thereby offers a rapid and reliable assay system for testing novel inhibitors of this system. Since L-S1 contains stably integrated human DNA in its genome, we postulate that the observed 5-HT-uptake system resulted from the expression of human gene(s) coding for the 5-HT transporter. Thus, cell lines such as L-S1 may represent novel means for screening and developing therapeutic agents specific for neurotransmitter-uptake systems as well as substrates for the cloning and elucidation of the genes encoding the various neurotransmitter transporters.Keywords
This publication has 30 references indexed in Scilit:
- The genomic clone G-21 which resembles a β-adrenergic receptor sequence encodes the 5-HT1A receptorNature, 1988
- Ion Dependence of Neurotransmitter Uptake: Inhibitory Effects of Ion SubstitutesJournal of Neurochemistry, 1987
- Sequence and functional expression of the GABAA receptor shows a ligand-gated receptor super-familyNature, 1987
- 5-Hydroxytryptamine Uptake and Imipramine Binding Sites in Neurotumor NCB-20 CellsJournal of Neurochemistry, 1985
- Analysis of pre- and postsynaptic factors of the serotonin system in rabbit retina.The Journal of cell biology, 1985
- Primary structures of β- and δ-subunit precursors of Torpedo californica acetylcholine receptor deduced from cDNA sequencesNature, 1983
- 2-Nitroimipramine: A selective irreversible inhibitor of [3H] serotonin uptake and [3H] imipramine binding in plateletsBiochemical and Biophysical Research Communications, 1981
- Demonstration of specific “high affinity” binding sites for [3H] imipramine on human plateletsLife Sciences, 1980
- THE UPTAKE AND RELEASE OF [3H]DOPAMINE IN THE GOLDFISH RETINAJournal of Neurochemistry, 1979
- Detection of specific sequences among DNA fragments separated by gel electrophoresisJournal of Molecular Biology, 1975