Automated phenotyping and lifespan assessment of a C. elegans model of Parkinson’s disease
Phenotypic analysis of Caenorhabditis elegans has greatly advanced our understanding of the molecular mechanisms implicated in the aging process as well as in age-related pathologies. However, conventional high-resolution imaging methods and survival assays are labor-intensive and subject to operato...
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doaj-c7fa509e3c144d7289007929cbe1face2021-04-02T16:34:44ZengKeAi Communications Co., Ltd.Translational Medicine of Aging2468-50112020-01-0143844Automated phenotyping and lifespan assessment of a C. elegans model of Parkinson’s diseaseMinwook Kim0Daniela Knoefler1Ellen Quarles2Ursula Jakob3Daphne Bazopoulou4Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, 48109, USADepartment of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, 48109, USA; Department of Biological Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA; Corresponding author. Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, 48109, USA.Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, 48109, USA; Corresponding author.Phenotypic analysis of Caenorhabditis elegans has greatly advanced our understanding of the molecular mechanisms implicated in the aging process as well as in age-related pathologies. However, conventional high-resolution imaging methods and survival assays are labor-intensive and subject to operator-based variations and decreased reproducibility. Recent advances in microfluidics and automated flatbed scanner technologies have significantly improved experimentation by eliminating handling errors and increasing the sensitivity in measurements. Here, we introduce a medium-throughput microfluidic platform, which efficiently positions and immobilizes single worms through pressurization for high resolution imaging. Worms are sorted based on select imaging criteria, and subsequently transferred into multi-well plates for automated lifespan assessment. To illustrate the applicability of this method, we imaged α-synuclein deposits in a C. elegans model of Parkinson’s Disease (PD). We found that age synchronized individuals expressing human α-synuclein vary greatly in the quantity and size of intracellular α-synuclein foci at early stages in life. Subsequent lifespan analysis of the individuals, however, did not reveal any correlation between the number or extent of α-synuclein deposits and subsequent lifespan. These studies suggest that the observed natural variations in α-synuclein deposits found in C. elegans models of PD do not originate from inherent differences in the fitness of the organism or contribute to alterations in lifespan.http://www.sciencedirect.com/science/article/pii/S2468501120300031C. elegansMicrofluidicsLifespanα-synuclein |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Minwook Kim Daniela Knoefler Ellen Quarles Ursula Jakob Daphne Bazopoulou |
spellingShingle |
Minwook Kim Daniela Knoefler Ellen Quarles Ursula Jakob Daphne Bazopoulou Automated phenotyping and lifespan assessment of a C. elegans model of Parkinson’s disease Translational Medicine of Aging C. elegans Microfluidics Lifespan α-synuclein |
author_facet |
Minwook Kim Daniela Knoefler Ellen Quarles Ursula Jakob Daphne Bazopoulou |
author_sort |
Minwook Kim |
title |
Automated phenotyping and lifespan assessment of a C. elegans model of Parkinson’s disease |
title_short |
Automated phenotyping and lifespan assessment of a C. elegans model of Parkinson’s disease |
title_full |
Automated phenotyping and lifespan assessment of a C. elegans model of Parkinson’s disease |
title_fullStr |
Automated phenotyping and lifespan assessment of a C. elegans model of Parkinson’s disease |
title_full_unstemmed |
Automated phenotyping and lifespan assessment of a C. elegans model of Parkinson’s disease |
title_sort |
automated phenotyping and lifespan assessment of a c. elegans model of parkinson’s disease |
publisher |
KeAi Communications Co., Ltd. |
series |
Translational Medicine of Aging |
issn |
2468-5011 |
publishDate |
2020-01-01 |
description |
Phenotypic analysis of Caenorhabditis elegans has greatly advanced our understanding of the molecular mechanisms implicated in the aging process as well as in age-related pathologies. However, conventional high-resolution imaging methods and survival assays are labor-intensive and subject to operator-based variations and decreased reproducibility. Recent advances in microfluidics and automated flatbed scanner technologies have significantly improved experimentation by eliminating handling errors and increasing the sensitivity in measurements. Here, we introduce a medium-throughput microfluidic platform, which efficiently positions and immobilizes single worms through pressurization for high resolution imaging. Worms are sorted based on select imaging criteria, and subsequently transferred into multi-well plates for automated lifespan assessment. To illustrate the applicability of this method, we imaged α-synuclein deposits in a C. elegans model of Parkinson’s Disease (PD). We found that age synchronized individuals expressing human α-synuclein vary greatly in the quantity and size of intracellular α-synuclein foci at early stages in life. Subsequent lifespan analysis of the individuals, however, did not reveal any correlation between the number or extent of α-synuclein deposits and subsequent lifespan. These studies suggest that the observed natural variations in α-synuclein deposits found in C. elegans models of PD do not originate from inherent differences in the fitness of the organism or contribute to alterations in lifespan. |
topic |
C. elegans Microfluidics Lifespan α-synuclein |
url |
http://www.sciencedirect.com/science/article/pii/S2468501120300031 |
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