title: Flotillin-1 is essential for PKC-triggered endocytosis and membrane microdomain localization of DAT.
abstract: 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.
authors.affiliation: Department of Neurology, Columbia University, College of Physicians and Surgeons, New York, New York, USA.
authors.forename: M Laura
authors.initials: ML
authors.lastname: Cremona
authors.forename: Heinrich J G
authors.initials: HJ
authors.lastname: Matthies
authors.forename: Kelvin
authors.initials: K
authors.lastname: Pau
authors.forename: Erica
authors.initials: E
authors.lastname: Bowton
authors.forename: Nicole
authors.initials: N
authors.lastname: Speed
authors.forename: Brandon J
authors.initials: BJ
authors.lastname: Lute
authors.forename: Monique
authors.initials: M
authors.lastname: Anderson
authors.forename: Namita
authors.initials: N
authors.lastname: Sen
authors.forename: Sabrina D
authors.initials: SD
authors.lastname: Robertson
authors.forename: Roxanne A
authors.initials: RA
authors.lastname: Vaughan
authors.forename: James E
authors.initials: JE
authors.lastname: Rothman
authors.forename: Aurelio
authors.initials: A
authors.lastname: Galli
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Javitch
authors.forename: Ai
authors.initials: A
authors.lastname: Yamamoto
mesh_terms: D000818:Animals; D050483:Dopamine Plasma Membrane Transport Proteins / Q000378:metabolism*; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D027342:Excitatory Amino Acid Transporter 2 / Q000378:metabolism / Q000502:physiology; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D021962:Membrane Microdomains / Q000378:metabolism*; D008565:Membrane Proteins / Q000502:physiology*; D051379:Mice; D009457:Neuroglia / Q000378:metabolism; D011493:Protein Kinase C / Q000502:physiology*
chemical_list: D050483:Dopamine Plasma Membrane Transport Proteins; D027342:Excitatory Amino Acid Transporter 2; D008565:Membrane Proteins; C106354:flotillins; D011493:Protein Kinase C