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Characterization of a Genetically Reconstituted High-Affinity System for Serotonin Transport
Albert Shu-Sen Chang, Jerome Victor Frnka, Danong Chen and Dominic Man-Kit Lam
Proceedings of the National Academy of Sciences of the United States of America
Vol. 86, No. 23 (Dec. 1, 1989), pp. 9611-9615
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/35134
Page Count: 5
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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 μ M; 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.
Proceedings of the National Academy of Sciences of the United States of America © 1989 National Academy of Sciences