Antidepressant-induced switch of beta 1-adrenoceptor trafficking as a mechanism for drug action.
Reduction in surface beta(1)-adrenoceptor (beta1AR) density is thought to play a critical role in mediating the therapeutic long term effects of antidepressants. Since antidepressants are neither agonists nor antagonists for G protein-coupled receptors, receptor density must be regulated through processes independent of direct receptor activation. Endocytosis and recycling of the beta1AR fused to green fluorescent protein at its carboxyl-terminus (beta1AR-GFP) were analyzed by confocal fluorescence microscopy of live cells and complementary ligand binding studies. In stably transfected C6 glioblastoma cells, beta1AR-GFP displayed identical ligand-binding isotherms and adenylyl cyclase activation as native beta1AR. Upon exposure to isoproterenol, a fraction of beta1AR-GFP (10-15%) internalized rapidly and colocalized with endocytosed transferrin receptors in an early endosomal compartment in the perinuclear region. Chronic treatment with the tricyclic antidepressant desipramine (DMI) did not affect internalization characteristics of beta1AR-GFP when challenged with isoproterenol. However, internalized receptors were not able to recycle back to the cell surface in DMI-treated cells, whereas recycling of transferrin receptors was not affected. Endocytosed receptors were absent from structures that stained with fluorescently labeled dextran, and inhibition of lysosomal protease activity did not restore receptor recycling, indicating that beta1AR-GFP did not immediately enter the lysosomal compartment. The data suggest a new mode of drug action causing a "switch" of receptor fate from a fast recycling pathway to a slowly exchanging perinuclear compartment. Antidepressant-induced reduction of receptor surface expression may thus be caused by modulation of receptor trafficking routes.
Bürgi
Sibylle
S
Department of Pharmacology, University of Bern, Switzerland.
Baltensperger
Kurt
K
Honegger
Ulrich E
UE
D000818:Animals; D000928:Antidepressive Agents / Q000494:pharmacology*; D003891:Desipramine / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects; D051381:Rats; D018342:Receptors, Adrenergic, beta-1 / Q000032:analysis / Q000187:drug effects* / Q000378:metabolism; D011990:Receptors, Transferrin / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014407:Tumor Cells, Cultured
D000928:Antidepressive Agents; D018342:Receptors, Adrenergic, beta-1; D011990:Receptors, Transferrin; D011993:Recombinant Fusion Proteins; D003891:Desipramine