Stable expression of the human dopamine transporter in N27 cells as an in vitro model for dopamine cell trafficking and metabolism. (October 2021)

• Record Type: Journal Article
• Authors: Cagle, B.S.; Sturgeon, M.L.; O'Brien, J.B.; Wilkinson, J.C.; Cornell, R.A.; Roman, D.L.; Doorn, J.A.
• Is Part Of: Toxicology in vitro. Volume 76(2021)
• Publication Date: 2021-10
• Subjects: Dopamine -- Dopamine transporter -- Neurodegeneration -- Dopamine metabolism -- Dopaminergic model; Dopamine DA -- 3, 4-dihydroxyphenylacetaldehyde DOPAL -- 3, 4-dihydroxphenylethanol DOPET -- 3, 4-dihydroxyphenylacetic acid DOPAC -- Homovanillic acid HVA -- Dopamine transporter DAT -- Human dopamine transporter hDAT -- 1-methyl-4-phenylpyridinium MPTP/MPP+ -- 6-hydroxydopamine 6-OHDA -- 4-hydroxynonenal 4-HNE -- 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide MTT -- Parkinson's disease PD -- Attention – deficit hyperactivity disorder ADHD -- Vesicular monoamine transporter VMAT -- Monoamine oxidase MAO -- Aldehyde dehydrogenase ALDH -- Aldose reductase AR -- False fluorescent neurotransmitter 102 FFN102; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2021.105210
• Languages: English
• ISSNs: 0887-2333

Abstract

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. Graphical abstract: Unlabelled Image Highlights: 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.