Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris .

Autor:	Odetti LM; Cátedra de Toxicología, Farmacología y Bioquímica Legal, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Ciudad Universitaria, Paraje El Pozo S/N 3000, Santa Fe, Argentina.; Consejo Nacional de Investigaciones Científicas y Técnicas Godoy Cruz 2290 C1425FQB Ciudad Autónoma de Buenos Aires, Argentina., Paravani EV; Lab. de Química Ambiental, Cátedra de Química General e Inorgánica. Facultad de Ingeniería-UNER, Oro Verde, Entre Ríos, Argentina., Simoniello MF; Cátedra de Toxicología, Farmacología y Bioquímica Legal, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Ciudad Universitaria, Paraje El Pozo S/N 3000, Santa Fe, Argentina., Poletta GL; Cátedra de Toxicología, Farmacología y Bioquímica Legal, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Ciudad Universitaria, Paraje El Pozo S/N 3000, Santa Fe, Argentina.; Consejo Nacional de Investigaciones Científicas y Técnicas Godoy Cruz 2290 C1425FQB Ciudad Autónoma de Buenos Aires, Argentina.; Proyecto Yacaré-Lab. de Zoología Aplicada: Anexo Vertebrados, Facultad de Humanidades y Ciencias-Universidad Nacional del Litoral/Ministerio de Medio Ambiente, Av. Aristóbulo del Valle 8700 3000, Santa Fe, Argentina.
Jazyk:	angličtina
Zdroj:	Heliyon [Heliyon] 2021 Feb 13; Vol. 7 (2), pp. e06253. Date of Electronic Publication: 2021 Feb 13 (Print Publication: 2021).
DOI:	10.1016/j.heliyon.2021.e06253
Abstrakt:	The quantitative real-time polymerase chain reaction (qPCR) has been one of the most promising approaches to perform rapid and accurate quantification of DNA in various biological systems. The aim of this study was to standardized the qPCR technique for the analysis of important genes involved in the main routes of antioxidant defense against reactive oxygen species (catalase: cat and superoxide dismutase: sod ) and evaluate the stability of different reference genes in blood of Caiman latirostris hatchlings. The stability of the reference genes, β-actin , glyceraldehyde 3-phosphate dehydrogenase ( gapdh ) and ribosomal protein L8 ( rpl8 ) was determined using the comparative ΔCt, NormFinder, geNorm, BestKeeper and RefFinder. Then, cat and sod genes were normalized with each reference gene and their mRNA abundances were determined through the qPCR. Stability of genes was ranked through the different methods in the following order: β-actin , rpl8 and gapdh , under normal physiological conditions. The results reveal that cat and sod genes present a similar relative mRNA abundance with β-actin and rpl8 . This is the first report of the analysis of antioxidant mRNA as potential biomarkers of oxidative stress in blood for all crocodilians species. Besides, we determined the stability of different reference genes that can be used for normalization of mRNA abundance patterns in blood of C. latirostris, without the need to sacrifice the animals. Competing Interests: The authors declare no conflict of interest. (© 2021 The Authors.)
Databáze:	MEDLINE
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