Abstract
In the nematode Caenorhabditis elegans (C. elegans), monitoring the lactate/pyruvate ratio in cells helps to detect imbalances in age-related energy metabolism. Here, we describe a modified small-scale extraction in C. elegans and measurement of lactate and pyruvate concentrations using colorimetric assay kits. During sample extraction, protein precipitation is the most critical step for precise determination of intracellular metabolites in C. elegans. Moreover, improved sensitivity and accuracy of colorimetric assay kits contributed to measurements of metabolites in samples derived from small-scale extraction. Using these protocols, we recently detected a metabolic alteration that occurs during aging by the monitoring the lactate/pyruvate ratio in a long-lived mutant of the mammalian tumor suppressor p53 ortholog CEP-1 in C. elegans.
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References
Derry WB, Putzke AP, Rothman JH (2001) Caenorhabditis elegans p53: role in apoptosis, meiosis, and stress resistance. Science 294:591–595
Schumacher B, Hofmann K, Boulton S, Gartner A (2001) The C. elegans homolog of the p53 tumor suppressor is required for DNA damage-induced apoptosis. Curr Biol 11:1722–1727
Arum O, Johnson TE (2007) Reduced expression of the Caenorhabditis elegans p53 ortholog cep-1 results in increased longevity. J Gerontol 62A:951–959
Ventura N, Rea SL, Schiavi A, Torgovnick A, Testi R, Johnson TE (2009) p53/CEP-1 increases or decreases lifespan, depending on level of mitochondrial bioenergetic stress. Aging Cell 8:380–393
Yanase S, Suda H, Yasuda K, Ishii N (2017) Impaired p53/CEP-1 is associated with lifespan extension through an age-related imbalance in the energy metabolism of C. elegans. Genes Cells 22:1004–1010
Warburg O (1956) On the origin of cancer cells. Science 123:309–314
Matoba S, Kang J-G, Patino WD, Wragg A, Boehm M, Gavrilova O et al (2006) p53 regulates mitochondrial respiration. Science 312:1650–1653
Kondoh H, Lleonart ME, Gil J, Wang J, Degan P, Peters G et al (2005) Glycolytic enzymes can modulate cellular life span. Cancer Res 65:177–185
Contractor T, Harris CR (2012) p53 negatively regulates transcription of the pyruvate dehydrogenase kinase Pdk2. Cancer Res 72:560–567
Taylor KACC (1996) A simple colorimetric assay for muramic acid and lactic acid. Appl Biochem Biotechnol 56:49–58
Lewis JA, Fleming JT (1995) Basic culture methods. Methods Cell Biol 48:3–29
Hosono R (1978) Sterilization and growth inhibition of Caenorhabditis elegans by 5-fluorodeoxyuridine. Exp Gerontol 13:369–374
Mitchell DH, Stiles JW, Santelli J, Sanadi DR (1979) Synchronous growth and aging of Caenorhabditis elegans in the presence of fluorodeoxyuridine. J Gerontol 34:28–36
Marbach EP, Weil MH (1967) Rapid enzymatic measurement of blood lactate and pyruvate: use and significance of metaphosphoric acid as a common precipitant. Clin Chem 13:314–325
Fabian TJ, Johnson TE (1994) Production of age-synchronous mass cultures of Caenorhabditis elegans. J Gerontol 49:B145–B156
Senoo-Matsuda N, Yasuda K, Tsuda M, Ohkubo T, Yoshimura S, Nakazawa H, Hartman PS et al (2001) A defect in the cytochrome b large subunit in complex II causes both superoxide anion overproduction and abnormal energy metabolism in Caenorhabditis elegans. J Biol Chem 276:41553–41558
Barker SB, Summerson WH (1941) The colorimetric determination of lactic acid in biological material. J Biol Chem 138:535–554
Acknowledgments
This work was supported by a special research grant from Daito Bunka University to S.Y.
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Yanase, S., Yasuda, K., Ishii, N. (2019). Monitoring Age-Related Changes in the Lactate/Pyruvate Ratio Using a Colorimetric Assay in a C. elegans Model of Increased Life Span. In: Guest, P. (eds) Pre-Clinical Models. Methods in Molecular Biology, vol 1916. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8994-2_12
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DOI: https://doi.org/10.1007/978-1-4939-8994-2_12
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