Evidence map›Paper›PMID 23859623›Full record

ArticleACS chemical biology2013

New fluorescent substrate enables quantitative and high-throughput examination of vesicular monoamine transporter 2 (VMAT2).

Gang Hu, Adam Henke, Richard J Karpowicz, Mark S Sonders, Frances Farrimond, Robert Edwards, David Sulzer, Dalibor Sames

Abstract readEvaluation Study
In one paragraph

Article in ACS chemical biology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 42 papers.

0numbers the graph read from it
0cells of the map it votes in
42citing papers in PubMed
1.8field-weighted citation impact, top 16% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

42 citing papers in PubMed, 72 citations in OpenAlex.

  1. Article
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  8. Tricyclic and tetracyclic antidepressants upregulate VMAT2 activity and rescue disease-causing VMAT2 variants.Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology · 2024
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  13. High-speed imaging reveals the bimodal nature of dense core vesicle exocytosis.Proceedings of the National Academy of Sciences of the United States of America · 2023
    Article
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  15. Review
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  18. Stable expression of the human dopamine transporter in N27 cells as an in vitro model for dopamine cell trafficking and metabolism.Toxicology in vitro : an international journal published in association with BIBRA · 2021
    Article
  19. Review
  20. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors at 2 institutions in 1 country.

Gang HuDepartment of Chemistry, Columbia University , New York, New York 10027, United States.
Adam Henke
Richard J Karpowicz
Mark S Sonders
Frances Farrimond
Robert Edwards
David Sulzer
Dalibor Sames
Columbia University · USUniversity of California, San Francisco · US

Funding

The Presynaptic Vesicle Cycle for the Slow Acting Neurotransmitter SecretionP01DA010154 · NIDA · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI KREITZER, ANATOL, SULZER, DAVID · 1995 to 2016
$14.7M
The Dopamine Synapse and Associative LearningR01DA007418 · NIDA · COLUMBIA UNIVERSITY HEALTH SCIENCES · PI David Sulzer · 1999 to 2026
$9.4M
TRANSPORT OF NEUROTRANSMITTERS INTO SYNAPTIC VESICLESR01MH050712 · NIMH · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI EDWARDS, ROBERT H · 1993 to 2011
$3.8M
Transport of Neurotransmitter Into Synaptic VesiclesR37MH050712 · NIMH · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI EDWARDS, ROBERT H · 2012 to 2020
$3.5M
Design of Molecular Probes For Optical Imaging of Dopamine NeurotransmissionR01MH086545 · NIMH · COLUMBIA UNIV NEW YORK MORNINGSIDE · PI SAMES, DALIBOR · 2010 to 2013
$1.5M
AMPHETAMINE ACTION MESOLIMBIC DOPAMINE NEURONAL CULTURER29DA007418 · NIDA · COLUMBIA UNIV NEW YORK MORNINGSIDE · PI SULZER, DAVID · 1991 to 1995
–
AMPHETAMINE ACTION IN MIDBRAIN DOPAMINE NEURONAL CULTURER55DA007418 · NIDA · NEW YORK STATE PSYCHIATRIC INSTITUTE · PI SULZER, DAVID · 1996 to 1996
–
NIDA NIH HHS DA07418NIDA NIH HHS DA10154NIDA NIH HHS P01 DA010154NIDA NIH HHS R01 DA007418NIMH NIH HHS MH086545NIMH NIH HHS MH50712NIMH NIH HHS R01 MH050712NIMH NIH HHS R01 MH086545NIMH NIH HHS R37 MH050712
6 · The paper itself

Abstract

Vesicular monoamine transporter 2 (VMAT2) is an essential component of the monoaminergic neurotransmission system in the brain as it transports monoamine neurotransmitters from the neuronal cytosol into the synaptic vesicles and thus contributes to modulation of neurotransmitter release. Considering the continuing interest in VMAT2 as a drug target, as well as a target for the design of imaging probes, we have developed a fluorescent substrate well suited for the study of VMAT2 in cell culture. Herein, we report the synthesis and characterization of a new fluorescent probe, FFN206, as an excellent VMAT2 substrate capable of detecting VMAT2 activity in intact cells using fluorescence microscopy, with subcellular localization to VMAT2-expressing acidic compartments without apparent labeling of other organelles. VMAT2 activity can also be measured via microplate reader. The apparent Km of FFN206 at VMAT2 was found to be 1.16 ± 0.10 μM, similar to that of dopamine. We further report the development and validation of a cell-based fluorescence assay amenable to high-throughput screening (HTS) using VMAT2-transfected HEK cells (Z'-factor of 0.7-0.8), enabling rapid identification of VMAT2 inhibitors and measurement of their inhibition constants over a broad range of affinities. FFN206 thus represents a new tool for optical examination of VMAT2 function in cell culture.

Indexed as

Drug Evaluation, PreclinicalFluorescent DyesHEK293 CellsHigh-Throughput Screening AssaysHumansMicroscopy, FluorescenceSensitivity and SpecificitySmall Molecule LibrariesVesicular Monoamine Transport ProteinsFluorescent DyesSmall Molecule LibrariesVesicular Monoamine Transport Proteins

Identifiers

PMID23859623
PMCPMC4557792
OpenAlexW1984533082

What OpenQuestion holds

Textmetadata
LicenceTDM
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.