Bridging the phenotypic gap: real-time assessment of mitochondrial function and metabolism of the nematode Caenorhabditis elegans

Cristina Lagido, Jonathan Pettitt, Aileen Winifred Flett, L Anne Glover

Research output: Contribution to journalArticle

34 Citations (Scopus)
3 Downloads (Pure)

Abstract

Background
The ATP levels of an organism are an important physiological parameter that is affected by genetic make up, ageing, stress and disease.

Results
We have generated luminescent C. elegans through ubiquitous, constitutive expression of firefly luciferase, widely used for in vitro ATP determination. We hypothesise that whole animal luminescence reflects its intracellular ATP levels in vivo. To test this, we characterised the bioluminescence response of C. elegans during sublethal exposure to, and recovery from azide, a treatment that inhibits mitochondrial respiration reversibly, and causes ATP depletion. Consistent with our expectations, in vivo luminescence decreased with increasing sublethal azide levels, and recovered fully when worms were removed from azide. Firefly luciferase expression levels, stability and activity did not influence the final luminescence. Bioluminescence also reflected the lowered activity of the electron transport chain achieved with RNA interference (RNAi) of genes encoding respiratory chain components.

Conclusion
Results indicated that C. elegans luminescence reports on ATP levels in real-time. For the first time, we are able to directly assess the metabolism of a whole, living, multicellular organism by determination of the relative ATP levels. This will enable genetic analysis based on a readily quantifiable metabolic phenotype and will provide novel insights into mechanisms of fitness and disease that are likely to be of relevance for other organisms, as well as the worm.

Original languageEnglish
Article number7
Number of pages9
JournalBMC Physiology
Volume8
DOIs
Publication statusPublished - 2 Apr 2008

Fingerprint

Caenorhabditis elegans
Adenosine Triphosphate
Luminescence
Azides
Firefly Luciferases
Electron Transport
RNA Interference
Respiration
Phenotype
Genes

Cite this

Bridging the phenotypic gap : real-time assessment of mitochondrial function and metabolism of the nematode Caenorhabditis elegans. / Lagido, Cristina; Pettitt, Jonathan; Flett, Aileen Winifred; Glover, L Anne.

In: BMC Physiology, Vol. 8, 7, 02.04.2008.

Research output: Contribution to journalArticle

@article{8186268b693649a8bb6c865cce590145,
title = "Bridging the phenotypic gap: real-time assessment of mitochondrial function and metabolism of the nematode Caenorhabditis elegans",
abstract = "Background The ATP levels of an organism are an important physiological parameter that is affected by genetic make up, ageing, stress and disease. Results We have generated luminescent C. elegans through ubiquitous, constitutive expression of firefly luciferase, widely used for in vitro ATP determination. We hypothesise that whole animal luminescence reflects its intracellular ATP levels in vivo. To test this, we characterised the bioluminescence response of C. elegans during sublethal exposure to, and recovery from azide, a treatment that inhibits mitochondrial respiration reversibly, and causes ATP depletion. Consistent with our expectations, in vivo luminescence decreased with increasing sublethal azide levels, and recovered fully when worms were removed from azide. Firefly luciferase expression levels, stability and activity did not influence the final luminescence. Bioluminescence also reflected the lowered activity of the electron transport chain achieved with RNA interference (RNAi) of genes encoding respiratory chain components. Conclusion Results indicated that C. elegans luminescence reports on ATP levels in real-time. For the first time, we are able to directly assess the metabolism of a whole, living, multicellular organism by determination of the relative ATP levels. This will enable genetic analysis based on a readily quantifiable metabolic phenotype and will provide novel insights into mechanisms of fitness and disease that are likely to be of relevance for other organisms, as well as the worm.",
author = "Cristina Lagido and Jonathan Pettitt and Flett, {Aileen Winifred} and Glover, {L Anne}",
year = "2008",
month = "4",
day = "2",
doi = "10.1186/1472-6793-8-7",
language = "English",
volume = "8",
journal = "BMC Physiology",
issn = "1472-6793",
publisher = "BioMed Central",

}

TY - JOUR

T1 - Bridging the phenotypic gap

T2 - real-time assessment of mitochondrial function and metabolism of the nematode Caenorhabditis elegans

AU - Lagido, Cristina

AU - Pettitt, Jonathan

AU - Flett, Aileen Winifred

AU - Glover, L Anne

PY - 2008/4/2

Y1 - 2008/4/2

N2 - Background The ATP levels of an organism are an important physiological parameter that is affected by genetic make up, ageing, stress and disease. Results We have generated luminescent C. elegans through ubiquitous, constitutive expression of firefly luciferase, widely used for in vitro ATP determination. We hypothesise that whole animal luminescence reflects its intracellular ATP levels in vivo. To test this, we characterised the bioluminescence response of C. elegans during sublethal exposure to, and recovery from azide, a treatment that inhibits mitochondrial respiration reversibly, and causes ATP depletion. Consistent with our expectations, in vivo luminescence decreased with increasing sublethal azide levels, and recovered fully when worms were removed from azide. Firefly luciferase expression levels, stability and activity did not influence the final luminescence. Bioluminescence also reflected the lowered activity of the electron transport chain achieved with RNA interference (RNAi) of genes encoding respiratory chain components. Conclusion Results indicated that C. elegans luminescence reports on ATP levels in real-time. For the first time, we are able to directly assess the metabolism of a whole, living, multicellular organism by determination of the relative ATP levels. This will enable genetic analysis based on a readily quantifiable metabolic phenotype and will provide novel insights into mechanisms of fitness and disease that are likely to be of relevance for other organisms, as well as the worm.

AB - Background The ATP levels of an organism are an important physiological parameter that is affected by genetic make up, ageing, stress and disease. Results We have generated luminescent C. elegans through ubiquitous, constitutive expression of firefly luciferase, widely used for in vitro ATP determination. We hypothesise that whole animal luminescence reflects its intracellular ATP levels in vivo. To test this, we characterised the bioluminescence response of C. elegans during sublethal exposure to, and recovery from azide, a treatment that inhibits mitochondrial respiration reversibly, and causes ATP depletion. Consistent with our expectations, in vivo luminescence decreased with increasing sublethal azide levels, and recovered fully when worms were removed from azide. Firefly luciferase expression levels, stability and activity did not influence the final luminescence. Bioluminescence also reflected the lowered activity of the electron transport chain achieved with RNA interference (RNAi) of genes encoding respiratory chain components. Conclusion Results indicated that C. elegans luminescence reports on ATP levels in real-time. For the first time, we are able to directly assess the metabolism of a whole, living, multicellular organism by determination of the relative ATP levels. This will enable genetic analysis based on a readily quantifiable metabolic phenotype and will provide novel insights into mechanisms of fitness and disease that are likely to be of relevance for other organisms, as well as the worm.

U2 - 10.1186/1472-6793-8-7

DO - 10.1186/1472-6793-8-7

M3 - Article

VL - 8

JO - BMC Physiology

JF - BMC Physiology

SN - 1472-6793

M1 - 7

ER -