Nanoscopic dopamine transporter distribution and conformation are inversely regulated by excitatory drive and D2 autoreceptor activity

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt


  • Fulltext

    Forlagets udgivne version, 8,81 MB, PDF-dokument

The nanoscopic organization and regulation of individual molecular components in presynaptic varicosities of neurons releasing modulatory volume neurotransmitters like dopamine (DA) remain largely elusive. Here we show, by application of several super-resolution microscopy techniques to cultured neurons and mouse striatal slices, that the DA transporter (DAT), a key protein in varicosities of dopaminergic neurons, exists in the membrane in dynamic equilibrium between an inward-facing nanodomain-localized and outward-facing unclustered configuration. The balance between these configurations is inversely regulated by excitatory drive and DA D2 autoreceptor activation in a manner dependent on Ca2+ influx via N-type voltage-gated Ca2+ channels. The DAT nanodomains contain tens of transporters molecules and overlap with nanodomains of PIP2 (phosphatidylinositol-4,5-bisphosphate) but show little overlap with D2 autoreceptor, syntaxin-1, and clathrin nanodomains. The data reveal a mechanism for rapid alterations of nanoscopic DAT distribution and show a striking link of this to the conformational state of the transporter.

TidsskriftCell Reports
Udgave nummer13
Antal sider27
StatusUdgivet - 2022

Bibliografisk note

Funding Information:
We thank Ralph Götz for support with expansion microscopy and Dr. Emiliana Borrelli for providing tissue from D2R knockout mice. The work was supported by Lundbeck Foundation grants R266-2017-4331 (to U.G.), R276-2018-792 (to U.G.), R230-2016-3154 (to M.D.L.), R181-2014-3090 (to F.H.), R303-2018-3540 (to F.H.), and R231-2016-2481-5 (to A.T.S.); Independent Research Fund Denmark – Medical Sciences ( 7016-00325B to U.G.); and NIDA-Intramural Research Program Z1A DA000610 (to A.H.N. and D.A.G.).

Publisher Copyright:
© 2022

ID: 322283247