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Intracellular Assessment of ATP Levels in Caenorhabditis elegans   

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Abstract

Eukaryotic cells heavily depend on adenosine triphosphate (ATP) generated by oxidative phosphorylation (OXPHOS) within mitochondria. ATP is the major energy currency molecule, which fuels cell to carry out numerous processes, including growth, differentiation, transportation and cell death among others (Khakh and Burnstock, 2009). Therefore, ATP levels can serve as a metabolic gauge for cellular homeostasis and survival (Artal-Sanz and Tavernarakis, 2009; Gomes et al., 2011; Palikaras et al., 2015). In this protocol, we describe a method for the determination of intracellular ATP levels using a bioluminescence approach in the nematode Caenorhabditis elegans.

Keywords: Ageing, ATP, Caenorhabditis elegans, Energy homeostasis, Luciferase, Metabolism, Mitochondria

Background

Mitochondria-derived ATP plays a crucial role in a variety of cellular and metabolic processes. Therefore, several methods have been developed to measure the levels of this important metabolite (Drew and Leeuwenburgh, 2003; Khan, 2003; Lagido et al., 2001; Vives-Bauza et al., 2007). In 1947, McElroy proposed the use of firefly bioluminescence to determine intracellular ATP levels, when he uncovered the essential role of ATP in light production (McElroy, 1947). The development of cloning and recombinant protein technology facilitated the development of the firefly luciferase assay to determine ATP levels (de Wet et al., 1985). Firefly luciferase, a monomeric 61 kD enzyme, catalyzes the oxidation of luciferin, which emits light at 560 nm. When ATP is the limiting factor of the enzymatic reaction, the intensity of light is proportional to the concentration of ATP. Thus, the use of a luminometer permits the detection of light intensity and subsequently the determination of ATP levels. In this protocol, we describe a method for ATP quantification using a bioluminescence approach in C. elegans.

Copyright: © 2016 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: Palikaras, K. and Tavernarakis, N. (2016). Intracellular Assessment of ATP Levels in Caenorhabditis elegans. Bio-protocol 6(23): e2048. DOI: 10.21769/BioProtoc.2048.
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