Stress and timing associated with Caenorhabditis elegans immobilization methods

Background: Caenorhabditis elegans is a model organism used to study gene, protein, and cell influence on function and behavior. These studies frequently require C. elegans to be immobilized for imaging or laser ablation experiments. There are a number of known techniques for immobilizing worms, but...

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Main Authors: Jacob R. Manjarrez, Roger Mailler
Format: Article
Language:English
Published: Elsevier 2020-07-01
Series:Heliyon
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2405844020311075
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author Jacob R. Manjarrez
Roger Mailler
author_facet Jacob R. Manjarrez
Roger Mailler
author_sort Jacob R. Manjarrez
collection DOAJ
description Background: Caenorhabditis elegans is a model organism used to study gene, protein, and cell influence on function and behavior. These studies frequently require C. elegans to be immobilized for imaging or laser ablation experiments. There are a number of known techniques for immobilizing worms, but to our knowledge, there are no comprehensive studies of the various agents in common use today. New method: This study determines the relationship between concentration, immobilization time, exposure time, and recovery likelihood for several immobilization agents. The agents used in this study are 1-Phenoxy-2-propanol, levamisole, sodium azide, polystyrene beads, and environmental cold shock. These tests are conducted using a humidified chamber to keep chemical concentrations consistent. Each of these agents is also tested to determine if they exhibit stress-related after effects using the gcs-1, daf-16, hsp-4, hif-1, hsp-16.2, and tmem-135 stress reporters. Results: We present a range of quick mount immobilization and recovery conditions for each agent tested. This study shows that, under controlled conditions, 1-Phenoxy-2-propanol shows significant stress from the daf-16 reporter. While 1-Phenoxy-2-propanol and sodium azide both create stress related after effects with long term recovery in the case of the hsp-16.2 reporter. Comparison with existing method(s): This study shows that commonly used concentrations of immobilizing agents are ineffective when evaporation is prevented. Conclusions: To improve reproducibility of results it is essential to use consistent concentrations of immobilizing agents. It is also critically important to account for stress-related after effects elicited by immobilization agents when designing any experiment.
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spelling doaj.art-eb34649e589f44e8b682363164bddf752022-12-21T18:42:26ZengElsevierHeliyon2405-84402020-07-0167e04263Stress and timing associated with Caenorhabditis elegans immobilization methodsJacob R. Manjarrez0Roger Mailler1University of Tulsa, 800 S. Tucker Dr., Tulsa, OK, 74104, USACorresponding author.; University of Tulsa, 800 S. Tucker Dr., Tulsa, OK, 74104, USABackground: Caenorhabditis elegans is a model organism used to study gene, protein, and cell influence on function and behavior. These studies frequently require C. elegans to be immobilized for imaging or laser ablation experiments. There are a number of known techniques for immobilizing worms, but to our knowledge, there are no comprehensive studies of the various agents in common use today. New method: This study determines the relationship between concentration, immobilization time, exposure time, and recovery likelihood for several immobilization agents. The agents used in this study are 1-Phenoxy-2-propanol, levamisole, sodium azide, polystyrene beads, and environmental cold shock. These tests are conducted using a humidified chamber to keep chemical concentrations consistent. Each of these agents is also tested to determine if they exhibit stress-related after effects using the gcs-1, daf-16, hsp-4, hif-1, hsp-16.2, and tmem-135 stress reporters. Results: We present a range of quick mount immobilization and recovery conditions for each agent tested. This study shows that, under controlled conditions, 1-Phenoxy-2-propanol shows significant stress from the daf-16 reporter. While 1-Phenoxy-2-propanol and sodium azide both create stress related after effects with long term recovery in the case of the hsp-16.2 reporter. Comparison with existing method(s): This study shows that commonly used concentrations of immobilizing agents are ineffective when evaporation is prevented. Conclusions: To improve reproducibility of results it is essential to use consistent concentrations of immobilizing agents. It is also critically important to account for stress-related after effects elicited by immobilization agents when designing any experiment.http://www.sciencedirect.com/science/article/pii/S2405844020311075GeneticsGene expressionGene regulationPromoterDose-response relationshipOxidative stress
spellingShingle Jacob R. Manjarrez
Roger Mailler
Stress and timing associated with Caenorhabditis elegans immobilization methods
Heliyon
Genetics
Gene expression
Gene regulation
Promoter
Dose-response relationship
Oxidative stress
title Stress and timing associated with Caenorhabditis elegans immobilization methods
title_full Stress and timing associated with Caenorhabditis elegans immobilization methods
title_fullStr Stress and timing associated with Caenorhabditis elegans immobilization methods
title_full_unstemmed Stress and timing associated with Caenorhabditis elegans immobilization methods
title_short Stress and timing associated with Caenorhabditis elegans immobilization methods
title_sort stress and timing associated with caenorhabditis elegans immobilization methods
topic Genetics
Gene expression
Gene regulation
Promoter
Dose-response relationship
Oxidative stress
url http://www.sciencedirect.com/science/article/pii/S2405844020311075
work_keys_str_mv AT jacobrmanjarrez stressandtimingassociatedwithcaenorhabditiselegansimmobilizationmethods
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