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The majority of drug screening assays for SLCs are based on the uptake of radiolabeled or fluorescent substrates. Thus, these approaches often have limitations that compromise on throughput or the physiological environment of the SLC. In this study, we report a novel application of an impedance-based biosensor, xCELLigence, to investigate dopamine transporter (DAT) activity via substrate-induced activation of G protein-coupled receptors (GPCRs). The resulting assay, which is coined the \u2018transporter activity through receptor activation\u2019 (TRACT) assay, is based on the hypothesis that DAT-mediated removal of extracellular dopamine directly affects the ability of dopamine to activate cognate membrane-bound GPCRs. In two human cell lines with heterologous DAT expression, dopamine-induced GPCR signaling was attenuated. Pharmacological inhibition or the absence of DAT restored the apparent potency of dopamine for GPCR activation. The inhibitory potencies for DAT inhibitors GBR12909 (pIC<jats:sub>50<\/jats:sub>\u2009=\u20096.2, 6.6) and cocaine (pIC<jats:sub>50<\/jats:sub>\u2009=\u20096.3) were in line with values from reported orthogonal transport assays. Conclusively, this study demonstrates the novel use of label-free whole-cell biosensors to investigate DAT activity using GPCR activation as a readout. This holds promise for other SLCs that share their substrate with a GPCR.<\/jats:p>","DOI":"10.1038\/s41598-020-79218-w","type":"journal-article","created":{"date-parts":[[2021,1,14]],"date-time":"2021-01-14T11:11:08Z","timestamp":1610622668000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["A study of the dopamine transporter using the TRACT assay, a novel in vitro tool for solute carrier drug discovery"],"prefix":"10.1038","volume":"11","author":[{"given":"Hubert J.","family":"Sijben","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie J. 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