5-Hydroxytryptamine Uptake and Imipramine Binding Sites in Neurotumor NCB-20 Cells

Abstract

NCB-20 cells (neuroblastoma × fetal Chinese hamster brain hybrids) are equipped with a [3H]5-hydroxytryptamine ([3H]5-HT) uptake system and [3H]imipramine recognition sites. Approximately 80% of the radioactivity taken up by cells incubated with [3H]5-HT was identified with 5-HT. [3H]5-HT uptake was temperature-dependent, partially sodium-dependent, saturable (Km= 7.3 ± 0.6 μM; Vmax= 2.0 ± 0.6 pmol/min/mg), and inhibited by clomipramine, imipramine, fluoxetine, and desipramine, but not by iprindole, mianserin, or opipramol. Lineweaver-Burk plots showed a competitive type of inhibition by imipramine and fluoxetine. [3H]5-HT uptake was not inhibited by nisoxetine or benztropine. [3H]Imipramine binding sites had a KD of 12 ± 2 nM and a Bmax of 22 ± 7 pmol/mg protein. The binding was sodium-sensitive although to a lesser extent than that found with brain membranes. Imipramine binding was displaced by tricyclic antidepressants with the following order of potency: clomipramine > imipramine > fluoxetine > desipramine ± iprindole = mianserin > opipramol. These results suggest that imipramine binding sites are present together with the 5-HT uptake sites in NCB-20 cells and that these sites interact functionally but are different biochemically.