The use of bioluminescent Caenorhabditis elegans as multicellular eukaryotic biosensors

Cristina Lagido, Jonathan Pettitt, Lesley Anne Glover

Research output: Contribution to conferenceAbstract

Abstract

Whole cell biosensors offer a powerful approach to environmental monitoring and toxicity testing, as they measure bioavailability of toxins and biological effects rather than total concentrations obtained by traditional analytical techniques. C. elegans is an excellent candidate for a biosensor that represents multicellular eukaryotes. We have constructed a strain of bioluminescent C. elegans with the firefly luciferase luc gene under the control of let-858 constitutive promoter. The transgenic nematodes luminesce after addition of the luciferin substrate and the amount of light emitted reflects the levels of intracellular ATP, providing a rapid, real-time indication of metabolic status. Conditions that affect their metabolism can be detected by a change in luminescence. Light reduction was observed in response to representative environmental insults such as increasing temperature, or concentrations of copper, lead and 3,5-DCP. This was due to a combination of increased mortality coupled with a decreased metabolic activity in the surviving animals. The luminescence assays are carried out in vivo and are rapid, sensitive and an easily quantifiable method for assessing toxicity. The assays lend themselves well to rapid screening and we are currently adapting them to a 96 well microplate format. Other potential applications of the technology developed here include development and screening of novel nematocide drugs and as a tool to study ageing.
Original languageEnglish
Publication statusPublished - 2000
EventEuropean Worm Meeting (2000) - Blankenberge, Belgium
Duration: 20 May 200023 May 2000

Conference

ConferenceEuropean Worm Meeting (2000)
CountryBelgium
CityBlankenberge
Period20/05/0023/05/00

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Biosensors
Toxicity
Luminescence
Antinematodal Agents
Assays
Screening
Biological Toxins
Firefly Luciferases
Metabolism
Copper
Animals
Genes
Adenosine Triphosphate
Aging of materials
Monitoring
Testing
Substrates
Pharmaceutical Preparations
Temperature
Biological Availability

Cite this

Lagido, C., Pettitt, J., & Glover, L. A. (2000). The use of bioluminescent Caenorhabditis elegans as multicellular eukaryotic biosensors. Abstract from European Worm Meeting (2000), Blankenberge, Belgium.

The use of bioluminescent Caenorhabditis elegans as multicellular eukaryotic biosensors. / Lagido, Cristina; Pettitt, Jonathan; Glover, Lesley Anne.

2000. Abstract from European Worm Meeting (2000), Blankenberge, Belgium.

Research output: Contribution to conferenceAbstract

Lagido, C, Pettitt, J & Glover, LA 2000, 'The use of bioluminescent Caenorhabditis elegans as multicellular eukaryotic biosensors' European Worm Meeting (2000), Blankenberge, Belgium, 20/05/00 - 23/05/00, .
Lagido C, Pettitt J, Glover LA. The use of bioluminescent Caenorhabditis elegans as multicellular eukaryotic biosensors. 2000. Abstract from European Worm Meeting (2000), Blankenberge, Belgium.
Lagido, Cristina ; Pettitt, Jonathan ; Glover, Lesley Anne. / The use of bioluminescent Caenorhabditis elegans as multicellular eukaryotic biosensors. Abstract from European Worm Meeting (2000), Blankenberge, Belgium.
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AU - Lagido, Cristina

AU - Pettitt, Jonathan

AU - Glover, Lesley Anne

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PY - 2000

Y1 - 2000

N2 - Whole cell biosensors offer a powerful approach to environmental monitoring and toxicity testing, as they measure bioavailability of toxins and biological effects rather than total concentrations obtained by traditional analytical techniques. C. elegans is an excellent candidate for a biosensor that represents multicellular eukaryotes. We have constructed a strain of bioluminescent C. elegans with the firefly luciferase luc gene under the control of let-858 constitutive promoter. The transgenic nematodes luminesce after addition of the luciferin substrate and the amount of light emitted reflects the levels of intracellular ATP, providing a rapid, real-time indication of metabolic status. Conditions that affect their metabolism can be detected by a change in luminescence. Light reduction was observed in response to representative environmental insults such as increasing temperature, or concentrations of copper, lead and 3,5-DCP. This was due to a combination of increased mortality coupled with a decreased metabolic activity in the surviving animals. The luminescence assays are carried out in vivo and are rapid, sensitive and an easily quantifiable method for assessing toxicity. The assays lend themselves well to rapid screening and we are currently adapting them to a 96 well microplate format. Other potential applications of the technology developed here include development and screening of novel nematocide drugs and as a tool to study ageing.

AB - Whole cell biosensors offer a powerful approach to environmental monitoring and toxicity testing, as they measure bioavailability of toxins and biological effects rather than total concentrations obtained by traditional analytical techniques. C. elegans is an excellent candidate for a biosensor that represents multicellular eukaryotes. We have constructed a strain of bioluminescent C. elegans with the firefly luciferase luc gene under the control of let-858 constitutive promoter. The transgenic nematodes luminesce after addition of the luciferin substrate and the amount of light emitted reflects the levels of intracellular ATP, providing a rapid, real-time indication of metabolic status. Conditions that affect their metabolism can be detected by a change in luminescence. Light reduction was observed in response to representative environmental insults such as increasing temperature, or concentrations of copper, lead and 3,5-DCP. This was due to a combination of increased mortality coupled with a decreased metabolic activity in the surviving animals. The luminescence assays are carried out in vivo and are rapid, sensitive and an easily quantifiable method for assessing toxicity. The assays lend themselves well to rapid screening and we are currently adapting them to a 96 well microplate format. Other potential applications of the technology developed here include development and screening of novel nematocide drugs and as a tool to study ageing.

M3 - Abstract

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