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Detection of Caenorhabditis elegans Germ Cell Apoptosis Following Exposure to Environmental Contaminant Mixtures: A Crude Oil-Dispersant Mixture Example.
Methods Mol Biol. 2021; 2326:3-18.MM

Abstract

Crude oil disasters, such as the Deepwater Horizon accident, have caused severe environmental contamination and damage, affecting the health of marine and terrestrial organisms. Some previous studies have demonstrated cleanup efforts using chemical dispersant induced more potent toxicities than oil alone due to an increase in bioavailability of crude oil components, such as PAHs. However, there still lacks a systematic procedure that provides methods to determine genotypic and phenotypic changes following exposure to environmental toxicants or toxicant mixture, such as dispersed crude oil. Here, we describe methods for identifying a mechanism of dispersed crude oil-induced reproductive toxicity in the model organisms, Caenorhabditis elegans (C. elegans). Due to the genetic malleability of C. elegans, two mutant strains outlined in this chapter were used to identify a pathway responsible for inducing apoptosis: MD701 bcIs39 [lim-7p::ced-1::GFP + lin-15(+)], a mutant strain that allows visualization of apoptotic bodies via a green fluorescent protein fused to CED-1; and TJ1 (cep-1(gk138) I.), a p53/CEP-1 defective strain that is unable to activate apoptosis via the p53/CEP-1 pathway. In addition, qRT-PCR was utilized to demonstrate the aberrant expression of apoptosis (ced-13, ced-3, ced-4, ced-9, cep-1, dpl-1, efl-1, efl-2, egl-1, egl-38, lin-35, pax-2, and sir-2.1) and cytochrome P450 (cyp14a3, cyp35a1, cyp35a2, cyp35a5, and cyp35c1) protein-coding genes following exposure to dispersed crude oil. The procedure outlined here can be applicable to determine whether environmental contaminants, most of time contaminant mixture, cause reproductive toxicity by activation of the proapoptotic, p53/CEP-1 pathway.

Authors+Show Affiliations

Department of Biology, East Carolina University, Greenville, NC, USA.No affiliation info availableDepartment of Biology, East Carolina University, Greenville, NC, USA. Panx@ecu.edu.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

34097257

Citation

Polli, Joseph Ryan, et al. "Detection of Caenorhabditis Elegans Germ Cell Apoptosis Following Exposure to Environmental Contaminant Mixtures: a Crude Oil-Dispersant Mixture Example." Methods in Molecular Biology (Clifton, N.J.), vol. 2326, 2021, pp. 3-18.
Polli JR, Farwell M, Pan X. Detection of Caenorhabditis elegans Germ Cell Apoptosis Following Exposure to Environmental Contaminant Mixtures: A Crude Oil-Dispersant Mixture Example. Methods Mol Biol. 2021;2326:3-18.
Polli, J. R., Farwell, M., & Pan, X. (2021). Detection of Caenorhabditis elegans Germ Cell Apoptosis Following Exposure to Environmental Contaminant Mixtures: A Crude Oil-Dispersant Mixture Example. Methods in Molecular Biology (Clifton, N.J.), 2326, 3-18. https://doi.org/10.1007/978-1-0716-1514-0_1
Polli JR, Farwell M, Pan X. Detection of Caenorhabditis Elegans Germ Cell Apoptosis Following Exposure to Environmental Contaminant Mixtures: a Crude Oil-Dispersant Mixture Example. Methods Mol Biol. 2021;2326:3-18. PubMed PMID: 34097257.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of Caenorhabditis elegans Germ Cell Apoptosis Following Exposure to Environmental Contaminant Mixtures: A Crude Oil-Dispersant Mixture Example. AU - Polli,Joseph Ryan, AU - Farwell,Mary, AU - Pan,Xiaoping, PY - 2021/6/7/entrez PY - 2021/6/8/pubmed PY - 2021/8/14/medline KW - Apoptosis KW - C. elegans KW - Environmental contaminants KW - GFP KW - Genotoxicity KW - Mixture toxicity KW - Oil spill KW - PCR SP - 3 EP - 18 JF - Methods in molecular biology (Clifton, N.J.) JO - Methods Mol Biol VL - 2326 N2 - Crude oil disasters, such as the Deepwater Horizon accident, have caused severe environmental contamination and damage, affecting the health of marine and terrestrial organisms. Some previous studies have demonstrated cleanup efforts using chemical dispersant induced more potent toxicities than oil alone due to an increase in bioavailability of crude oil components, such as PAHs. However, there still lacks a systematic procedure that provides methods to determine genotypic and phenotypic changes following exposure to environmental toxicants or toxicant mixture, such as dispersed crude oil. Here, we describe methods for identifying a mechanism of dispersed crude oil-induced reproductive toxicity in the model organisms, Caenorhabditis elegans (C. elegans). Due to the genetic malleability of C. elegans, two mutant strains outlined in this chapter were used to identify a pathway responsible for inducing apoptosis: MD701 bcIs39 [lim-7p::ced-1::GFP + lin-15(+)], a mutant strain that allows visualization of apoptotic bodies via a green fluorescent protein fused to CED-1; and TJ1 (cep-1(gk138) I.), a p53/CEP-1 defective strain that is unable to activate apoptosis via the p53/CEP-1 pathway. In addition, qRT-PCR was utilized to demonstrate the aberrant expression of apoptosis (ced-13, ced-3, ced-4, ced-9, cep-1, dpl-1, efl-1, efl-2, egl-1, egl-38, lin-35, pax-2, and sir-2.1) and cytochrome P450 (cyp14a3, cyp35a1, cyp35a2, cyp35a5, and cyp35c1) protein-coding genes following exposure to dispersed crude oil. The procedure outlined here can be applicable to determine whether environmental contaminants, most of time contaminant mixture, cause reproductive toxicity by activation of the proapoptotic, p53/CEP-1 pathway. SN - 1940-6029 UR - https://www.unboundmedicine.com/medline/citation/34097257/Detection_of_Caenorhabditis_elegans_Germ_Cell_Apoptosis_Following_Exposure_to_Environmental_Contaminant_Mixtures:_A_Crude_Oil_Dispersant_Mixture_Example_ L2 - https://dx.doi.org/10.1007/978-1-0716-1514-0_1 DB - PRIME DP - Unbound Medicine ER -