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

Lycas, Matthew D.; Ejdrup, Aske L; Sørensen, Andreas T.; Haahr, Nicolai O.; Jørgensen, S.; Guthrie, Daryl A.; Støier, Jonatan Fullerton; Werner, Christian; Newman, Amy Hauck; Sauer, Markus; Herborg, Freja; Gether, Ulrik

doi:10.1016/j.celrep.2022.111431

Cited by 15 publications

(38 citation statements)

References 82 publications

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“…We observed no significant redistribution of Stx1 after treatment (Fig. S3c), matching previous findings when analyzed with tessellation 30 . But tessellation may not uncover all reorganizational nuances.…”

Section: Assessing Colocalization Of Synaptic Release Proteinssupporting

confidence: 91%

“…The two transporters mediate clearance of their respective substrates, dopamine and noradrenaline, and play an important role in shaping the spatiotemporal profile of dopaminergic and noradrenergic neurotransmission 31,32 . Interestingly, DAT and NET display differences in subcellular localization and trafficking itinerary 30,[33][34][35] . Specifically, NET has been reported to internalize more rapidly than DAT and preferentially sort to the Rab11 recycling compartment rather than for degradation, whereas the internalized DAT sort less to this compartment.…”

Section: Investigating Protein Trafficking Itinerariesmentioning

confidence: 99%

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A density-based enrichment measure for assessing colocalization in single-molecule localization microscopy data

et al. 2022

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Dual-color single-molecule localization microscopy (SMLM) provides unprecedented possibilities for detailed studies of colocalization of different molecular species in a cell. However, the informational richness of the data is not fully exploited by current analysis tools that often reduce colocalization to a single value. Here, we describe a tool specifically designed for determination of co-localization in both 2D and 3D from SMLM data. The approach uses a function that describes the relative enrichment of one molecular species on the density distribution of a reference species. The function reframes the question of colocalization by providing a density-context relevant to multiple biological questions. Moreover, the function visualize enrichment (i.e. colocalization) directly in the images for easy interpretation. We demonstrate the approach’s functionality on both simulated data and cultured neurons, and compare it to current alternative measures. The method is available in a Python function for easy and parameter-free implementation.

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Section: Assessing Colocalization Of Synaptic Release Proteinssupporting

confidence: 91%

Section: Investigating Protein Trafficking Itinerariesmentioning

confidence: 99%

A density-based enrichment measure for assessing colocalization in single-molecule localization microscopy data

et al. 2022

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“…Novel super-resolution imaging has shown that DAT is sequestered into nanodomains in the plasma membrane to allow for highly regulated control of DA dynamics via DAT distribution. , Further localization to these nanoclusters may denote DAT conformations, as nanoclustered DAT proteins were found to be in the inward-facing conformation . Interestingly, the D2R antagonism decreased localization of DAT to nanodomains, thereby promoting the unclustering of DAT proteins which are, in contrast, in outward-facing conformation . Thus, the blunting of D2R feedback, like that seen at times of high estradiol levels in females, , may act synergistically with the cocaine/typical DAT inhibitors to increase unclustering of DAT proteins.…”

Section: Resultsmentioning

confidence: 99%

“…We postulate that the lack of sex-dependent differences following the administration of atypical DAT inhibitors like R- modafinil could support the differences between typical and atypical DAT inhibitors in altering the DAT number or function in the neuronal membrane. Novel super-resolution imaging has shown that DAT is sequestered into nanodomains in the plasma membrane to allow for highly regulated control of DA dynamics via DAT distribution. , Further localization to these nanoclusters may denote DAT conformations, as nanoclustered DAT proteins were found to be in the inward-facing conformation . Interestingly, the D2R antagonism decreased localization of DAT to nanodomains, thereby promoting the unclustering of DAT proteins which are, in contrast, in outward-facing conformation .…”

Section: Resultsmentioning

confidence: 99%

An FSCV Study on the Effects of Targeted Typical and Atypical DAT Inhibition on Dopamine Dynamics in the Nucleus Accumbens Shell of Male and Female Mice

Hersey

Chen

Bartole

et al. 2023

ACS Chem. Neurosci.

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Understanding the neurochemistry underlying sex differences in psychostimulant use disorders (PSUD) is essential for developing related therapeutics. Many psychostimulants, like cocaine, inhibit the dopamine transporter (DAT), which is largely thought to account for actions related to their misuse and dependence. Cocaine-like, typical DAT inhibitors preferentially bind DAT in an outward-facing conformation, while atypical DAT inhibitors, like modafinil, prefer a more inward-facing DAT conformation. Modafinil and R-modafinil have emerged as potential therapeutic options for selected populations of individuals affected by PSUD. In addition, analogs of modafinil (JJC8-088 and JJC8-091) with different pharmacological profiles have been explored as potential PSUD medications in preclinical models. In this work, we employ fast scan cyclic voltammetry (FSCV) to probe nucleus accumbens shell (NAS) dopamine (DA) dynamics in C57BL/6 male and female mice. We find that cocaine slowed DA clearance in both male and female mice but produced more robust increases in evoked NAS DA in female mice. R-Modafinil produced mild increases in evoked NAS DA and slowed DA clearance across the sexes. The modafinil analog JJC8-088, a typical DAT inhibitor, produced increases in evoked NAS DA in female and male mice. Finally, JJC8-091, an atypical DAT inhibitor, produced limited increases in evoked NAS DA and slowed DA clearance in both sexes. In this work we begin to tease out how sex differences may alter the effects of DAT targeting and highlight how this may help focus research toward effective treatment options for PSUD.

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“…Of the four tested compounds, DG3‐80 and DG4‐91 demonstrated specific surface labelling of DAT in both systems. DG3‐80 has recently been used for d STORM experiments revealing a mechanism for rapid alterations in nanoscopic DAT distribution, demonstrating a remarkable link to the conformational state of the transporter in both cultured neurons and mouse striatal slices 50 …”

Section: Matsmentioning

confidence: 99%

Illuminating the monoamine transporters: Fluorescently labelled ligands to study dopamine, serotonin and norepinephrine transporters

Camacho‐Hernandez

Jahan

Newman

2023

Basic Clin Pharma Tox

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Fluorescence microscopy has revolutionized the visualization of physiological processes in live‐cell systems. With the recent innovations in super resolution microscopy, these events can be examined with high precision and accuracy. The development of fluorescently labelled small molecules has provided a significant advance in understanding the physiological relevance of targeted proteins that can now be visualized at the cellular level. One set of physiologically important target proteins are the monoamine transporters (MATs) that play an instrumental role in maintaining monoamine signalling homeostasis. Understanding the mechanisms underlying their regulation and dysregulation is fundamental to treating several neuropsychiatric conditions such as attention deficit hyperactivity disorder (ADHD), anxiety, depression and substance use disorders. Herein, we describe the rationale behind the small molecule design of fluorescently labelled ligands (FLL) either as MAT substrates or inhibitors as well as their applications to advance our understanding of this class of transporters in health and disease.

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

Cited by 15 publications

References 82 publications

A density-based enrichment measure for assessing colocalization in single-molecule localization microscopy data

A density-based enrichment measure for assessing colocalization in single-molecule localization microscopy data

An FSCV Study on the Effects of Targeted Typical and Atypical DAT Inhibition on Dopamine Dynamics in the Nucleus Accumbens Shell of Male and Female Mice

Illuminating the monoamine transporters: Fluorescently labelled ligands to study dopamine, serotonin and norepinephrine transporters

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