Abstract
To date, most in vitro toxicity testing has focused on acute effects of compounds at high concentrations. This testing strategy does not reflect real-life exposures, which might contribute to long-term disease outcome. We used a 3D-human dopaminergic in vitro LUHMES cell line model to determine whether effects of short-term rotenone exposure (100 nM, 24 h) are permanent or reversible. A decrease in complex I activity, ATP, mitochondrial diameter, and neurite outgrowth were observed acutely. After compound removal, complex I activity was still inhibited; however, ATP levels were increased, cells were electrically active and aggregates restored neurite outgrowth integrity and mitochondrial morphology. We identified significant transcriptomic changes after 24 h which were not present 7 days after wash-out. Our results suggest that testing short-term exposures in vitro may capture many acute effects which cells can overcome, missing adaptive processes, and long-term mechanisms. In addition, to study cellular resilience, cells were re-exposed to rotenone after wash-out and recovery period. Pre-exposed cells maintained higher metabolic activity than controls and presented a different expression pattern in genes previously shown to be altered by rotenone. NEF2L2, ATF4, and EAAC1 were downregulated upon single hit on day 14, but unchanged in pre-exposed aggregates. DAT and CASP3 were only altered after re-exposure to rotenone, while TYMS and MLF1IP were downregulated in both single-exposed and pre-exposed aggregates. In summary, our study shows that a human cell-based 3D model can be used to assess cellular adaptation, resilience, and long-term mechanisms relevant to neurodegenerative research.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1-20 |
| Number of pages | 20 |
| Journal | Archives of Toxicology |
| DOIs | |
| State | Accepted/In press - Jun 28 2018 |
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Keywords
- 3D LUHMES
- Cellular defence
- Gene response
- Neurodegeneration
- Pesticide
- Recovery
- Resilience
- Rotenone
ASJC Scopus subject areas
- Toxicology
- Health, Toxicology and Mutagenesis
Cite this
Toxicity, recovery, and resilience in a 3D dopaminergic neuronal in vitro model exposed to rotenone. / Harris, Georgina; Eschment, Melanie; Orozco, Sebastian Perez; McCaffery, J. Michael; Maclennan, Richard; Severin, Daniel; Leist, Marcel; Kleensang, Andre; Pamies, David; Maertens, Alexandra; Hogberg, Helena; Freeman, Dana; Kirkwood, Alfredo; Hartung, Thomas; Smirnova, Lena.
In: Archives of Toxicology, 28.06.2018, p. 1-20.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Toxicity, recovery, and resilience in a 3D dopaminergic neuronal in vitro model exposed to rotenone
AU - Harris, Georgina
AU - Eschment, Melanie
AU - Orozco, Sebastian Perez
AU - McCaffery, J. Michael
AU - Maclennan, Richard
AU - Severin, Daniel
AU - Leist, Marcel
AU - Kleensang, Andre
AU - Pamies, David
AU - Maertens, Alexandra
AU - Hogberg, Helena
AU - Freeman, Dana
AU - Kirkwood, Alfredo
AU - Hartung, Thomas
AU - Smirnova, Lena
PY - 2018/6/28
Y1 - 2018/6/28
N2 - To date, most in vitro toxicity testing has focused on acute effects of compounds at high concentrations. This testing strategy does not reflect real-life exposures, which might contribute to long-term disease outcome. We used a 3D-human dopaminergic in vitro LUHMES cell line model to determine whether effects of short-term rotenone exposure (100 nM, 24 h) are permanent or reversible. A decrease in complex I activity, ATP, mitochondrial diameter, and neurite outgrowth were observed acutely. After compound removal, complex I activity was still inhibited; however, ATP levels were increased, cells were electrically active and aggregates restored neurite outgrowth integrity and mitochondrial morphology. We identified significant transcriptomic changes after 24 h which were not present 7 days after wash-out. Our results suggest that testing short-term exposures in vitro may capture many acute effects which cells can overcome, missing adaptive processes, and long-term mechanisms. In addition, to study cellular resilience, cells were re-exposed to rotenone after wash-out and recovery period. Pre-exposed cells maintained higher metabolic activity than controls and presented a different expression pattern in genes previously shown to be altered by rotenone. NEF2L2, ATF4, and EAAC1 were downregulated upon single hit on day 14, but unchanged in pre-exposed aggregates. DAT and CASP3 were only altered after re-exposure to rotenone, while TYMS and MLF1IP were downregulated in both single-exposed and pre-exposed aggregates. In summary, our study shows that a human cell-based 3D model can be used to assess cellular adaptation, resilience, and long-term mechanisms relevant to neurodegenerative research.
AB - To date, most in vitro toxicity testing has focused on acute effects of compounds at high concentrations. This testing strategy does not reflect real-life exposures, which might contribute to long-term disease outcome. We used a 3D-human dopaminergic in vitro LUHMES cell line model to determine whether effects of short-term rotenone exposure (100 nM, 24 h) are permanent or reversible. A decrease in complex I activity, ATP, mitochondrial diameter, and neurite outgrowth were observed acutely. After compound removal, complex I activity was still inhibited; however, ATP levels were increased, cells were electrically active and aggregates restored neurite outgrowth integrity and mitochondrial morphology. We identified significant transcriptomic changes after 24 h which were not present 7 days after wash-out. Our results suggest that testing short-term exposures in vitro may capture many acute effects which cells can overcome, missing adaptive processes, and long-term mechanisms. In addition, to study cellular resilience, cells were re-exposed to rotenone after wash-out and recovery period. Pre-exposed cells maintained higher metabolic activity than controls and presented a different expression pattern in genes previously shown to be altered by rotenone. NEF2L2, ATF4, and EAAC1 were downregulated upon single hit on day 14, but unchanged in pre-exposed aggregates. DAT and CASP3 were only altered after re-exposure to rotenone, while TYMS and MLF1IP were downregulated in both single-exposed and pre-exposed aggregates. In summary, our study shows that a human cell-based 3D model can be used to assess cellular adaptation, resilience, and long-term mechanisms relevant to neurodegenerative research.
KW - 3D LUHMES
KW - Cellular defence
KW - Gene response
KW - Neurodegeneration
KW - Pesticide
KW - Recovery
KW - Resilience
KW - Rotenone
UR - http://www.scopus.com/inward/record.url?scp=85049143580&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85049143580&partnerID=8YFLogxK
U2 - 10.1007/s00204-018-2250-8
DO - 10.1007/s00204-018-2250-8
M3 - Article
C2 - 29955902
AN - SCOPUS:85049143580
SP - 1
EP - 20
JO - Archiv fur Toxikologie
JF - Archiv fur Toxikologie
SN - 0003-9446
ER -