Journal article
Stable expression of the human dopamine transporter in N27 cells as an in vitro model for dopamine cell trafficking and metabolism
Toxicology in vitro, Vol.76, p.105210
07/2021
DOI: 10.1016/j.tiv.2021.105210
PMCID: PMC8419135
PMID: 34252731
Abstract
Dopamine (DA) metabolism and cell trafficking are critical for the proper functioning of DA neurons. Disruption of these DA processes can yield toxic products and is implicated in neurological conditions including Parkinson's disease (PD). To investigate pathogenic mechanisms involving DA neurons, in vitro models that recapitulate DA metabolism and trafficking in vivo are crucial. N27 cells are a widely used model for PD; however, these cells exhibit little expression of the DA transporter (DAT) confounding studies of DA uptake and metabolism. This lack of adequate DAT expression calls into question the use of this cell line as a model to study DA cell trafficking and metabolism. To overcome this problem, we stably expressed the human DAT (hDAT) in N27 cells to develop cells that we named N27-BCD. This approach allows for characterization of toxicants that may alter DA metabolism, trafficking, and/or interactions with DAT. N27-BCD cells are more sensitive to the neurotoxins 1-methyl-4-phenylpyridinium (MPTP/MPP+) and 6-hydroxydopamine (6-OHDA). N27-BCD cells allowed for clear observation of DA metabolism, whereas N27 cells did not. Here, we propose that stable expression of hDAT in N27 cells yields a useful model of DA neurons to study the impact of altered DA cell trafficking and metabolism.
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•hDAT-N27 cells (N27-BCD) provide a model of dopamine metabolism and trafficking.•hDAT expression yields measurable dopamine metabolism via HPLC-PDA detection.•N27 cells expressing hDAT are more sensitive to DAT-dependent neurotoxicants.
Details
- Title: Subtitle
- Stable expression of the human dopamine transporter in N27 cells as an in vitro model for dopamine cell trafficking and metabolism
- Creators
- B S Cagle - Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, University of Iowa, 180 S Grand Ave. Iowa City, Iowa 52242, USAM.L Sturgeon - The Interdisciplinary Graduate Program in Molecular Medicine, Carver College of Medicine, University of Iowa, 451 Newton Road, Iowa City, Iowa 52242, USAJ.B O'Brien - Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, University of Iowa, 180 S Grand Ave. Iowa City, Iowa 52242, USAJ.C Wilkinson - Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, University of Iowa, 180 S Grand Ave. Iowa City, Iowa 52242, USAR.A Cornell - Department of Anatomy and Cell Biology, Carver College of Medicine, University of Iowa, 51 Newton Road Iowa City, Iowa 52242, USAD.L Roman - Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, University of Iowa, 180 S Grand Ave. Iowa City, Iowa 52242, USAJ.A Doorn - Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, University of Iowa, 180 S Grand Ave. Iowa City, Iowa 52242, USA
- Resource Type
- Journal article
- Publication Details
- Toxicology in vitro, Vol.76, p.105210
- DOI
- 10.1016/j.tiv.2021.105210
- PMID
- 34252731
- PMCID
- PMC8419135
- NLM abbreviation
- Toxicol In Vitro
- ISSN
- 0887-2333
- eISSN
- 1879-3177
- Publisher
- Elsevier Ltd
- Language
- English
- Date published
- 07/2021
- Academic Unit
- Pharmacy; Anatomy and Cell Biology; Iowa Neuroscience Institute; Pharmaceutical Sciences and Experimental Therapeutics; Dental Research; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984102307402771
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