Chapter 29
Evaluation of Carotenoids Protection Against Oxidative
Stress in the Animal Model Caenorhabditis elegans
Patricia Martorell, Silvia Llopis, Jose ´ Vicente Gil, Salvador Genove ´ s,
Daniel Ramo ´ n, Lorenzo Zacarı ´as, and Marı ´a Jesu ´ s Rodrigo
Abstract
The nematode Caenorhabditis elegans is a versatile and powerful model organism for animal experimental
research and, despite being an invertebrate, displays remarkably similar molecular bases and conserved
cellular pathways to those of humans. Oxidative stress is an etiological factor that influences numerous
diseases, degenerative processes and aging. C. elegans has revealed as an opportune and feasible organism to
investigate the antioxidant effects of different bioactives or complex food matrices, and a number of
protocols have been developed by using different oxidative stressors. Carotenoids are recognized as
quenchers and scavengers of reactive oxygen species, and many of their related health benefits attributed
in the diet are tightly linked to their antioxidant properties. In this chapter, we report a simple and rapid
assay to evaluate the protection capacity of pure carotenoids or complex carotenoid extracts against
oxidative stress in the model system C. elegans. The protocol describes a representative feeding experiment
by adding carotenoids to the nematode growth medium and after an incubation period, the C. elegans
populations fed with carotenoids are exposed to an acute oxidative stress by using H 2 O 2 as oxidative agent.
The protection against oxidative stress is evaluated as the survival rate of the nematodes fed with the
carotenoid prior to receiving oxidative treatment compared with the survival rate of control nematode
population. In order to confirm the carotenoid intake by the nematodes during the feeding experiment a
bioassimilation experiment is also reported.
Key words Antioxidant assay, Bioactive ingredients, Caenorhabditis elegans, Carotenoids, Experimental animal model, Oxidative stress
1 Introduction
The free-living soil nematode Caenorhabditis elegans was adopted
in the 1960s as model organism for developmental biology [1] and
since then its use as experimental system for many human pathological disorders and as a useful platform to investigate food ingredient or drug bioactive effects, has been increasing and becoming
more relevant [2]. C. elegans offers many advantages for laboratory
manipulation such as small size (1 mm) and transparent body,
Manuel Rodrı ´guez-Concepcio ´ n and Ralf Welsch (eds.), Plant and Food Carotenoids: Methods and Protocols,
Methods in Molecular Biology, vol. 2083, https://doi.org/10.1007/978-1-4939-9952-1_29,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
387
Evaluation of Carotenoids Protection Against Oxidative
Stress in the Animal Model Caenorhabditis elegans
Patricia Martorell, Silvia Llopis, Jose ´ Vicente Gil, Salvador Genove ´ s,
Daniel Ramo ´ n, Lorenzo Zacarı ´as, and Marı ´a Jesu ´ s Rodrigo
Abstract
The nematode Caenorhabditis elegans is a versatile and powerful model organism for animal experimental
research and, despite being an invertebrate, displays remarkably similar molecular bases and conserved
cellular pathways to those of humans. Oxidative stress is an etiological factor that influences numerous
diseases, degenerative processes and aging. C. elegans has revealed as an opportune and feasible organism to
investigate the antioxidant effects of different bioactives or complex food matrices, and a number of
protocols have been developed by using different oxidative stressors. Carotenoids are recognized as
quenchers and scavengers of reactive oxygen species, and many of their related health benefits attributed
in the diet are tightly linked to their antioxidant properties. In this chapter, we report a simple and rapid
assay to evaluate the protection capacity of pure carotenoids or complex carotenoid extracts against
oxidative stress in the model system C. elegans. The protocol describes a representative feeding experiment
by adding carotenoids to the nematode growth medium and after an incubation period, the C. elegans
populations fed with carotenoids are exposed to an acute oxidative stress by using H 2 O 2 as oxidative agent.
The protection against oxidative stress is evaluated as the survival rate of the nematodes fed with the
carotenoid prior to receiving oxidative treatment compared with the survival rate of control nematode
population. In order to confirm the carotenoid intake by the nematodes during the feeding experiment a
bioassimilation experiment is also reported.
Key words Antioxidant assay, Bioactive ingredients, Caenorhabditis elegans, Carotenoids, Experimental animal model, Oxidative stress
1 Introduction
The free-living soil nematode Caenorhabditis elegans was adopted
in the 1960s as model organism for developmental biology [1] and
since then its use as experimental system for many human pathological disorders and as a useful platform to investigate food ingredient or drug bioactive effects, has been increasing and becoming
more relevant [2]. C. elegans offers many advantages for laboratory
manipulation such as small size (1 mm) and transparent body,
Manuel Rodrı ´guez-Concepcio ´ n and Ralf Welsch (eds.), Plant and Food Carotenoids: Methods and Protocols,
Methods in Molecular Biology, vol. 2083, https://doi.org/10.1007/978-1-4939-9952-1_29,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
387