Optimization of experimental and computational processes to study mitochondrial trafficking and metabolic biomarkers
(2018) 52nd Annual Meeting of the European Society for Clinical Investigation In European Journal of Clinical Investigation 48(51). p.89-89- Abstract
- Background: The complex polarity of neurons requires specialized mechanisms to allocate sufficient number of mitochondria to neurites and synapses to control local calcium and ATP levels. Deficits in trafficking are directly linked to neurodegeneration. Our objective is to demonstrate that alterations of mitochondrial spatial location and metabolism are interconnected and can lead to point-of-no-return situations characterized as the first signs of metabolic rupture.
Material and methods: The neuroblastoma cell line SH-SY5Y was used and differentiated using different protocols to develop a neuronal phenotype. To disturb mitochondrial metabolism and traffic, two mitochondrial poisons, rotenone and 6-hydroxydopamine, were used.... (More) - Background: The complex polarity of neurons requires specialized mechanisms to allocate sufficient number of mitochondria to neurites and synapses to control local calcium and ATP levels. Deficits in trafficking are directly linked to neurodegeneration. Our objective is to demonstrate that alterations of mitochondrial spatial location and metabolism are interconnected and can lead to point-of-no-return situations characterized as the first signs of metabolic rupture.
Material and methods: The neuroblastoma cell line SH-SY5Y was used and differentiated using different protocols to develop a neuronal phenotype. To disturb mitochondrial metabolism and traffic, two mitochondrial poisons, rotenone and 6-hydroxydopamine, were used. Cells were labelled with different fluorescent probes and imaged under a INCell Analyzer. Cell mass and metabolic activity were measured using the SRB and resazurin assays, respectively.
Results: We have shown that a seven-day cell differentiation protocol using retinoic acid provided cells with better neuronal-like morphology, which are suited to study mitochondrial metabolic and dynamic parameters. We also demonstrate that rotenone and 6-hydroxydopamine toxicity is time and dose dependent, altering cell and mitochondria morphology, as well as mitochondrial membrane potential.
Conclusions: Images obtained with increasing concentrations of mitochondrial toxicants showed a gradual effect on different mitochondrial parameters that can be quantified using computational tools. These effects will be paired with metabolic data obtained for the same drug concentrations at later timepoints, to assess the consequences of mitochondrial disruption on cell viability. Building on the collected experimental information, we are training machine learning classification algorithms to accurately predict point-of-no-return situations that unbalance the cell to a state of metabolic catastrophe. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/2d85fe39-f314-428c-8242-0830b7f3bebe
- author
- Simoes, Rui LU ; Cunha-Oliveira, Teresa ; Oliveira, Paulo and Pereira, Francisco
- publishing date
- 2018
- type
- Contribution to journal
- publication status
- published
- in
- European Journal of Clinical Investigation
- volume
- 48
- issue
- 51
- article number
- P045-T
- pages
- 89 - 89
- publisher
- Wiley-Blackwell
- conference name
- 52nd Annual Meeting of the European Society for Clinical Investigation
- conference location
- Barcelona, Spain
- conference dates
- 2018-05-30 - 2018-06-01
- ISSN
- 0014-2972
- DOI
- 10.1111/eci.12926
- language
- English
- LU publication?
- no
- id
- 2d85fe39-f314-428c-8242-0830b7f3bebe
- date added to LUP
- 2021-09-21 20:23:49
- date last changed
- 2021-09-23 11:33:01
@misc{2d85fe39-f314-428c-8242-0830b7f3bebe,
abstract = {{Background: The complex polarity of neurons requires specialized mechanisms to allocate sufficient number of mitochondria to neurites and synapses to control local calcium and ATP levels. Deficits in trafficking are directly linked to neurodegeneration. Our objective is to demonstrate that alterations of mitochondrial spatial location and metabolism are interconnected and can lead to point-of-no-return situations characterized as the first signs of metabolic rupture.<br/><br/>Material and methods: The neuroblastoma cell line SH-SY5Y was used and differentiated using different protocols to develop a neuronal phenotype. To disturb mitochondrial metabolism and traffic, two mitochondrial poisons, rotenone and 6-hydroxydopamine, were used. Cells were labelled with different fluorescent probes and imaged under a INCell Analyzer. Cell mass and metabolic activity were measured using the SRB and resazurin assays, respectively.<br/><br/>Results: We have shown that a seven-day cell differentiation protocol using retinoic acid provided cells with better neuronal-like morphology, which are suited to study mitochondrial metabolic and dynamic parameters. We also demonstrate that rotenone and 6-hydroxydopamine toxicity is time and dose dependent, altering cell and mitochondria morphology, as well as mitochondrial membrane potential.<br/><br/>Conclusions: Images obtained with increasing concentrations of mitochondrial toxicants showed a gradual effect on different mitochondrial parameters that can be quantified using computational tools. These effects will be paired with metabolic data obtained for the same drug concentrations at later timepoints, to assess the consequences of mitochondrial disruption on cell viability. Building on the collected experimental information, we are training machine learning classification algorithms to accurately predict point-of-no-return situations that unbalance the cell to a state of metabolic catastrophe.}},
author = {{Simoes, Rui and Cunha-Oliveira, Teresa and Oliveira, Paulo and Pereira, Francisco}},
issn = {{0014-2972}},
language = {{eng}},
note = {{Conference Abstract}},
number = {{51}},
pages = {{89--89}},
publisher = {{Wiley-Blackwell}},
series = {{European Journal of Clinical Investigation}},
title = {{Optimization of experimental and computational processes to study mitochondrial trafficking and metabolic biomarkers}},
url = {{http://dx.doi.org/10.1111/eci.12926}},
doi = {{10.1111/eci.12926}},
volume = {{48}},
year = {{2018}},
}