{"doi":"10.1016/j.jneumeth.2012.06.002","title":"A fluorescent-based assay for live cell, spatially resolved assessment of vesicular monoamine transporter 2-mediated neurotransmitter transport","abstract":null,"journal":"Journal of Neuroscience Methods","year":2012,"id":593095,"datarank":1.0236601001015915,"base_score":3.332204510175204,"endowment":3.332204510175204,"self_citation_contribution":0.49983067652628066,"citation_network_contribution":0.5238294235753108,"self_endowment_contribution":0.49983067652628066,"citer_contribution":0.5238294235753108,"corpus_percentile":null,"corpus_rank":null,"citation_count":27,"citer_count":18,"citers_with_citation_signal":15,"citers_with_endowment":15,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":598777,"name":"Kristen A. Stout","orcid":"0000-0002-3641-5326","position":1,"is_corresponding":false},{"id":87740,"name":"Gary W. Miller","orcid":"0000-0001-8984-1284","position":2,"is_corresponding":false},{"id":456639,"name":"Alison I. Bernstein","orcid":"0000-0002-5589-4318","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"A fluorescent-based assay for live cell, spatially resolved assessment of vesicular monoamine transporter 2-mediated neurotransmitter transport","abstract":"The vesicular monoamine transporter 2 (VMAT2; Slc18a2) packages monoamines into synaptic vesicles. Monoamine homeostasis is highly regulated and dysfunction may play a role in Parkinson's disease, Huntington's disease, drug addiction, and neuropsychiatric disorders. The primary function of VMAT2 is to sequester monoamine neurotransmitters into vesicles for subsequent release; it also sequesters toxicants away from cytosolic sites of action. Identification of compounds that modify the action of VMAT2 may be useful as therapeutic agents for preventing or reversing monoamine-related toxicity. Current methods for measuring VMAT2 function are unable to assess uptake in intact cells. Here, we adapted the Neurotransmitter Uptake Assay (Molecular Devices) to develop a measure of VMAT2 function in live whole cells. This assay contains a fluorescent compound, which is transported into cells by the plasma membrane monoamine transporters and has been marketed as a rapid, high-throughput, plate reader based assay for function of these plasma membrane transporters. We demonstrate a modified version of this assay that can be used to visualize and measure transport into vesicles by VMAT2. HEK293 cell lines stably expressing the dopamine transporter and a mCherry-VMAT2 fusion protein were generated. Confocal microscopy confirmed that the fluorescent compound is transported into mCherry-positive compartments. Furthermore, the VMAT2-specific inhibitor tetrabenazine (TBZ) blocks uptake into the mCherry-positive compartment. Confocal images can be analyzed to generate a measure of VMAT2 activity. In summary, we demonstrate a method for spatially resolved analysis of VMAT2-mediated uptake in live intact cells.","is_dataset_classified":null,"base_score":0.0,"endowment":0.0,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"22698664","pmcid":"PMC3429701","openalex_id":null,"authors":[],"funders":[{"funder_name":"NIEHS NIH HHS","grant_id":"P01 ES016731","title":null},{"funder_name":"NIEHS NIH HHS","grant_id":"T32 ES012870","title":null},{"funder_name":"NINDS NIH HHS","grant_id":"P30 NS055077","title":null},{"funder_name":"NIA NIH HHS","grant_id":"P50 AG025688","title":null},{"funder_name":"NIDA NIH HHS","grant_id":"T32 DA015040","title":null}],"total_grants":5,"fwci":null,"citation_percentile":null,"influential_citations":0,"citation_trend":[],"oa_status":null,"license":"https://www.elsevier.com/legal/tdmrep-license","oa_locations":[{"url":"https://api.elsevier.com/content/article/PII:S0165027012002142?httpAccept=text/xml","host_type":"publisher"},{"url":"https://api.elsevier.com/content/article/PII:S0165027012002142?httpAccept=text/plain","host_type":"publisher"}],"fields_of_study":[],"mesh_terms":["Humans","Tritium","Dopamine","Piperazines","Tetrabenazine","Luminescent Proteins","Adrenergic Uptake Inhibitors","Transfection","Gene Expression Regulation","Protein Transport","Dose-Response Relationship, Drug","Dopamine Plasma Membrane Transport Proteins","Vesicular Monoamine Transport Proteins","HEK293 Cells","Red Fluorescent Protein"],"keywords":[],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-26T17:11:21.537737Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}