RT Journal Article SR Electronic T1 Amphetamine Induces a Calcium/Calmodulin-Dependent Protein Kinase II-Dependent Reduction in Norepinephrine Transporter Surface Expression Linked to Changes in Syntaxin 1A/Transporter Complexes JF Molecular Pharmacology JO Mol Pharmacol FD American Society for Pharmacology and Experimental Therapeutics SP 230 OP 239 DO 10.1124/mol.106.026690 VO 71 IS 1 A1 Concetta Dipace A1 Uhna Sung A1 Francesca Binda A1 Randy D. Blakely A1 Aurelio Galli YR 2007 UL http://molpharm.aspetjournals.org/content/71/1/230.abstract AB Norepinephrine (NE) transporters (NETs) are high-affinity transport proteins that mediate the synaptic clearance of NE after vesicular release. NETs represent a major therapeutic target for antidepressants and are targets of multiple psychostimulants including amphetamine (AMPH) and cocaine. Recently, we demonstrated that syntaxin 1A (SYN1A) regulates NET surface expression and, through binding to the transporter's NH2 terminus, regulates transporter catalytic function. AMPH induces NE efflux and may also regulate transporter trafficking. We monitored NET distribution and function in catecholaminergic cell lines (CAD) stably transfected with either full-length human NET (CAD-hNET) or with an hNET N-terminal deletion (CAD-hNETΔ28-47 cells). In hNET-CAD cells, AMPH causes a slow and small reduction of surface hNET with a modest increase in hNET/SYN1A associations at the plasma membrane. In contrast, in CAD-hNETΔ28-47 cells, AMPH induces a rapid and substantial reduction in surface hNETΔ28-47 accompanied by a large increase in plasma membrane hNETΔ28-47/SYN1A complexes. We also found that AMPH in CAD-hNETΔ28-47 cells induces a robust increase in cytosolic Ca2+ and concomitant activation of calcium/calmodulin-dependent protein kinase II (CaMKII). Inhibition of either the increase in intracellular Ca2+ or CaMKII activity blocks AMPH-stimulated hNETΔ28-47 trafficking and the formation of hNETΔ28-47/SYN1A complexes. Here, we demonstrate that AMPH stimulation of CAMKII stabilizes an hNET/SYN1A complex. This hNET/SYN1A complex rapidly redistributes, upon AMPH treatment, when mechanisms supported by the transporter's NH2 terminus are eliminated. The American Society for Pharmacology and Experimental Therapeutics