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J. Biol. Chem., Vol. 269, Issue 44, 27351-27356, Nov, 1994
KJ Miller and BJ Hoffman
Many antidepressants inhibit 5-hydroxytryptamine (5HT) transport resulting
in increased 5HT levels in the synapse. However, physiological regulation
of neurotransmitter uptake has not been demonstrated. We have examined the
effect of receptor-activated second messengers on the 5HT transporter in
rat basophilic leukemia cells (RBL 2H3). Here, we show that activation of
an A3 adenosine receptor results in an increase of 5HT uptake in RBL cells,
due to an increase in maximum velocity (Vmax). The A3 adenosine
receptor-stimulated increase in transport is blocked by inhibitors of
nitric oxide synthase and by a cGMP-dependent kinase inhibitor. In fact,
compounds that generate nitric oxide (NO) and the cGMP analog 8-bromo-cGMP
mimicked the effect of A3 receptor stimulation, suggesting that the
elevation in transport occurs through the generation of the gaseous second
messenger NO and a subsequent elevation in cGMP. Additionally, the 5HT
transporter is differentially regulated by second messengers since direct
activation of protein kinase C by phorbol esters decreases 5HT uptake by
decreasing Vmax. Our results suggest that the changes in transport are due
to a direct modification of the 5HT transporter, possibly by
phosphorylation, which appears to alter the rate at which transport occurs.
As the 5HT transporter in RBL cells is identical to that in neurons, our
results suggest that analogous mechanisms may operate in the brain.
Adenosine A3 receptors regulate serotonin transport via nitric oxide and cGMP
Laboratory of Cell Biology, National Institute of Mental Health, Rockville Pike, Bethesda, Maryland 20892.
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