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Flotillin-1 is essential for PKC-triggered endocytosis and membrane microdomain localization of DAT

2011; Nature Portfolio; Volume: 14; Issue: 4 Linguagem: Inglês

10.1038/nn.2781

1546-1726

María Laura Cremona, H Matthies, Kelvin Pau, Erica Bowton, Nicole Speed, Brandon J. Lute, Monique Anderson, Namita Sen, Sabrina D. Robertson, Roxanne A. Vaughan, James E. Rothman, Aurelio Galli, Jonathan A. Javitch, Ai Yamamoto,

Ion channel regulation and function

The authors report that the protein Flotillin-1/Reggie-2 (Flot1) is required for PKC-regulated internalization of the dopamine transporter (DAT) and the glial glutamate transporter EAAT2. Flot1 was required to localize DAT within plasma membrane microdomains in stable cell lines, and for amphetamine-induced reverse transport of dopamine in neurons. Plasmalemmal neurotransmitter transporters (NTTs) regulate the level of neurotransmitters, such as dopamine (DA) and glutamate, after their release at brain synapses. Stimuli including protein kinase C (PKC) activation can lead to the internalization of some NTTs and a reduction in neurotransmitter clearance capacity. We found that the protein Flotillin-1 (Flot1), also known as Reggie-2, was required for PKC-regulated internalization of members of two different NTT families, the DA transporter (DAT) and the glial glutamate transporter EAAT2, and we identified a conserved serine residue in Flot1 that is essential for transporter internalization. Further analysis revealed that Flot1 was also required to localize DAT within plasma membrane microdomains in stable cell lines, and was essential for amphetamine-induced reverse transport of DA in neurons but not for DA uptake. In sum, our findings provide evidence for a critical role of Flot1-enriched membrane microdomains in PKC-triggered DAT endocytosis and the actions of amphetamine.