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Reference gene selection for qPCR in mussel, Mytilus edulis, during gametogenesis and exogenous estrogen exposure

Rotchell, Jeanette; Cubero-Leon, Elena; Ciocan, Corina M.; Minier, Christophe; Rotchell, Jeanette M.

Authors

Elena Cubero-Leon

Corina M. Ciocan

Christophe Minier



Abstract

Purpose: The aim of this study is to develop a normalization method for real-time PCR data by analyzing the most stably expressed control genes in mussel (Mytilus edulis) reproductive tissue. Methods: To facilitate this, six candidate genes, including several commonly used in the literature, were investigated in mussels at different stages of gametogenesis and following experimental exposure to a model estrogen (17b-estradiol). GeNorm and NormFinder softwares were employed to assess the stability of the reference genes. Results: Our results demonstrate that the most stable reference genes are not the same in mussels at different stages of gametogenesis and in experimentally E2-exposed mussels. Interestingly, HEL (helicase) and ACT (actin) mRNA expression levels were most affected by the stage of gametogenesis and yet, in molluscan studies, ACT is possibly the most frequently used reference gene. Conclusions: We demonstrate that the experimental results are highly dependent on the reference gene chosen and that statistically significant contrasting differences between sample groups are present or absent depending on the reference gene employed. © 2012 Springer-Verlag.

Journal Article Type Article
Publication Date 2012-08
Journal ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
Print ISSN 0944-1344
Electronic ISSN 1614-7499
Publisher Springer Verlag
Peer Reviewed Peer Reviewed
Volume 19
Issue 7
Pages 2728-2733
APA6 Citation Cubero-Leon, E., Ciocan, C. M., Minier, C., & Rotchell, J. M. (2012). Reference gene selection for qPCR in mussel, Mytilus edulis, during gametogenesis and exogenous estrogen exposure. Environmental science and pollution research international, 19(7), (2728-2733). doi:10.1007/s11356-012-0772-9. ISSN 0944-1344
DOI https://doi.org/10.1007/s11356-012-0772-9
Keywords Mytilus edulis; Reference genes; Normalization; Endocrine disruption; Real-time PCR; Toxicology