Quantum dots reveal heterogeneous membrane diffusivity and dynamic surface density polarization of dopamine transporter

English version

Autoři: Oleg Kovtun ^aff001; Ian D. Tomlinson ^aff001; Riley S. Ferguson ^aff001; Sandra J. Rosenthal ^aff001
Působiště autorů: Department of Chemistry, Vanderbilt University, Nashville, Tennessee, United States of America ^aff001; Vanderbilt Institute of Nanoscale Science and Engineering, Vanderbilt University, Nashville, Tennessee, United States of America ^aff002; Department of Pharmacology, Vanderbilt University, Nashville, Tennessee, United States of America ^aff003; Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, Tennessee, United States of America ^aff004; Department of Physics and Astronomy, Vanderbilt University, Nashville, Tennessee, United States of America ^aff005; Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, Tennessee, United States of America ^aff006
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pone.0225339

Souhrn

The presynaptic dopamine transporter mediates rapid reuptake of synaptic dopamine. Although cell surface DAT trafficking recently emerged as an important component of DAT regulation, it has not been systematically investigated. Here, we apply our single quantum dot (Qdot) tracking approach to monitor DAT plasma membrane dynamics in several heterologous expression cell hosts with nanometer localization accuracy. We demonstrate that Qdot-tagged DAT proteins exhibited highly heterogeneous membrane diffusivity dependent on the local membrane topography. We also show that Qdot-tagged DATs were localized away from the flat membrane regions and were dynamically retained in the membrane protrusions and cell edges for the duration of imaging. Single quantum dot tracking of wildtype DAT and its conformation-defective coding variants (R60A and W63A) revealed a significantly accelerated rate of dysfunctional DAT membrane diffusion. We believe our results warrant an in-depth investigation as to whether compromised membrane dynamics is a common feature of brain disorder-derived DAT mutants.

Klíčová slova:

Cell membranes – Membrane proteins – Cholesterol – Dopamine – Mass diffusivity – Cytoskeleton – Membrane trafficking – Dopamine transporters

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Quantum dots reveal heterogeneous membrane diffusivity and dynamic surface density polarization of dopamine transporter

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