Quantum dots reveal heterogeneous membrane diffusivity and dynamic surface density polarization of dopamine transporter
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 monito...
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| Published in | PloS one Vol. 14; no. 11; p. e0225339 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
Public Library of Science
21.11.2019
Public Library of Science (PLoS) |
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| Online Access | Get full text |
| ISSN | 1932-6203 1932-6203 |
| DOI | 10.1371/journal.pone.0225339 |
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| Abstract | 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. |
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| AbstractList | 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. 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.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. |
| Audience | Academic |
| Author | Ferguson, Riley S. Kovtun, Oleg Tomlinson, Ian D. Rosenthal, Sandra J. |
| AuthorAffiliation | Beijing Forestry University, CHINA 2 Vanderbilt Institute of Nanoscale Science and Engineering, Vanderbilt University, Nashville, Tennessee, United States of America 3 Department of Pharmacology, Vanderbilt University, Nashville, Tennessee, United States of America 4 Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, Tennessee, United States of America 5 Department of Physics and Astronomy, Vanderbilt University, Nashville, Tennessee, United States of America 6 Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, Tennessee, United States of America 1 Department of Chemistry, Vanderbilt University, Nashville, Tennessee, United States of America |
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| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/31751387$$D View this record in MEDLINE/PubMed |
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| SubjectTerms | Algorithms Amphetamines Animals Biology and Life Sciences Cell Membrane - chemistry Cell Membrane - metabolism Cell membranes Cell surface Chemistry Cholesterol Cocaine Coding Conformation Diffusion rate Diffusivity Dopamine Dopamine Plasma Membrane Transport Proteins - chemistry Dopamine Plasma Membrane Transport Proteins - metabolism Dopamine transporter Electronics Engineering HEK293 Cells Humans Kinases Localization Membranes Microscopy Models, Theoretical Mutants Neural coding Neuroimaging Phenols (Class of compounds) Physical Sciences Proteins Quantum Dots Reproducibility of Results Sperm Structure-Activity Relationship Tracking |
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| Title | Quantum dots reveal heterogeneous membrane diffusivity and dynamic surface density polarization of dopamine transporter |
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