Dataset Information

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.

SUBMITTER: Lagido C

PROVIDER: S-EPMC2364618 | biostudies-literature | 2008 Apr

REPOSITORIES: biostudies-literature

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Publications

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

Lagido Cristina C Pettitt Jonathan J Flett Aileen A Glover L Anne LA

BMC physiology 20080402

<h4>Background</h4>The ATP levels of an organism are an important physiological parameter that is affected by genetic make up, ageing, stress and disease.<h4>Results</h4>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 subl ...[more]

PMID: 18384668

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Project description:Introduction:Biomedical and translational research often relies on the evaluation of patients or specimens that meet specific clinical or laboratory criteria. The typical approach used to identify biospecimens is a manual, retrospective process that exists outside the clinical workflow. This often makes biospecimen collection cost prohibitive and prevents the collection of analytes with short stability times. Emerging data architectures offer novel approaches to enhance specimen-identification practices. To this end, we present a new tool that can be deployed in a real-time environment to automate the identification and notification of available biospecimens for biomedical research. Methods:Real-time clinical and laboratory data from Cloverleaf (Infor, NY, NY) were acquired within our computational health platform, which is built on open-source applications. Study-specific filters were developed in NiFi (Apache Software Foundation, Wakefield, MA, USA) to identify the study-appropriate specimens in real time. Specimen metadata were stored in Elasticsearch (Elastic N. V., Mountain View, CA, USA) for visualization and automated alerting. Results:Between June 2018 and December 2018, we identified 2992 unique specimens belonging to 2815 unique patients, split between two different use cases. Based on laboratory policy for specimen retention and study-specific stability requirements, secure E-mail notifications were sent to investigators to automatically notify of availability. The assessment of throughput on commodity hardware demonstrates the ability to scale to approximately 2000 results per second. Conclusion:This work demonstrates that real-world clinical data can be analyzed in real time to increase the efficiency of biospecimen identification with minimal overhead for the clinical laboratory. Future work will integrate additional data types, including the analysis of unstructured data, to enable more complex cases and biospecimen identification.

Project description:Studies of the biological effects of low-level and below-background radiation are important in understanding the potential effects of radiation exposure in humans. To study this issue we exposed the nematode Caenorhabditis elegans to average background and below-background radiation levels. Two experiments were carried-out in the underground radiation biology laboratory at the Waste Isolation Pilot Plant (WIPP) in New Mexico USA. The first experiment used naïve nematodes with data collected within 1 week of being placed underground. The second experiment used worms that were incubated for 8 months underground at below background radiation levels. Nematode eggs were placed in two incubators, one at low radiation (ca.15.6 nGy/hr) and one supplemented with 2 kg of natural KCl (ca. 67.4 nGy/hr). Phenotypic variables measured were: (1) egg hatching success (2) body size from larval development to adulthood, (3) developmental time from egg to egg laying adult, and (4) egg laying rate of young adult worms. Transcriptome analysis was performed on the first experiment on 72 h old adult worms. Within 72 h of being underground, there was a trend of increased egg-laying rate in the below-background radiation treatment. This trend became statistically significant in the group of worms exposed to below-background radiation for 8 months. Worms raised for 8 months in these shielded conditions also had significantly faster growth rates during larval development. Transcriptome analyses of 72-h old naïve nematode RNA showed significant differential expression of genes coding for sperm-related proteins and collagen production. In the below-background radiation group, the genes for major sperm protein (msp, 42% of total genes) and sperm-related proteins (7.5%) represented 49.5% of the total genes significantly up-regulated, while the majority of down-regulated genes were collagen (col, 37%) or cuticle-related (28%) genes. RT-qPCR analysis of target genes confirmed transcriptomic data. These results demonstrate that exposure to below-background radiation rapidly induces phenotypic and transcriptomic changes in C. elegans within 72 h of being brought underground.

Dataset Information

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

Background

Results

Conclusion

Publications

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

Similar Datasets

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