Protein kinase C activation regulates human serotonin transporters in HEK-293 cells via altered cell surface expression.

Qian Y, Galli A, Ramamoorthy S, Risso S, DeFelice LJ, Blakely RD
J Neurosci. 1997 17 (1): 45-57

PMID: 8987735 · PMCID: PMC6793697

Antidepressant- and cocaine-sensitive serotonin (5-hydroxytryptamine, 5-HT) transporters (SERTs) dictate clearance of extracellular 5-HT after release. To explore protein kinase C-mediated SERT regulation, we generated a stable human SERT (hSERT)-expressing cell line (293-hSERT) and evaluated modulation of 5-HT activity via studies of 5-HT flux, hSERT-mediated currents under voltage clamp, and surface distribution of SERT protein. 293-hSERT cells exhibit saturable, high-affinity, and antidepressant-sensitive 5-HT uptake as well as hSERT-dependent whole-cell currents. In these cells, the protein kinase C activator beta-PMA caused a time-dependent reduction in 5-HT uptake capacity (Vmax) after acute application and a reduction in SERT-mediated currents. Effects of beta-PMA were mimicked by the phorbol ester beta-PDBu, were not observed with the inactive alpha-isomers, and could be blocked by treatment of cells with the protein kinase C inhibitor staurosporine. Biotinylation/immunoblot analyses showed that activity reductions are paralleled by a staurosporine-sensitive loss of surface SERT protein. These data indicate that altered surface abundance, rather than reduced catalytic transport efficiency, mediates acute PKC-dependent modulation of 5-HT uptake.

MeSH Terms (13)

Antidepressive Agents Carrier Proteins Cell Line Cell Membrane Electric Conductivity Enzyme Activation Humans Membrane Glycoproteins Membrane Transport Proteins Nerve Tissue Proteins Protein Kinase C Serotonin Serotonin Plasma Membrane Transport Proteins

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