A new set of endogenous control genes for use in quantitative real-time PCR experiments show that formin Ldia2dex transcripts are enriched in the early embryo of the pond snail Lymnaea stagnalis (Panpulmonata)

Johnson, Harriet F.; Davison, Angus

Abstract

© 2019 The Author(s). Published by Oxford University Press on behalf of The Malacological Society of London, all rights reserved. Although the pond snail Lymnaea stagnalis is an emerging model organism for molecular studies in a wide variety of fields, there are a limited number of verified endogenous control genes for use in quantitative real-Time PCR. As part of a larger study on snail chirality, or left-right asymmetry, we assayed gene expression in pond snail embryos. We evaluated six candidate control genes, by comparing their expression in three tissues (ovotestis, foot and embryo) and used three software programmes (geNorm, Normfinder and Bestkeeper) to do so. The specific utility of these control genes was then tested by investigating the relative expression of six experimental transcripts, including formin Ldia2, a gene that has been associated with chiral variation in L. stagnalis. All six control genes were found to be suitable for use in the three tissues tested. Of the six experimental genes, it was found that all were relatively depleted in the early embryo compared with other tissues, except the formin Ldia2 gene. Instead, transcripts of the wild-Type Ldia2dex were enriched in the embryo, whereas a nonfunctional frameshifted version, Ldia2sin, was severely depleted. These differences in Ldia2sin expression were less evident in the ovotestis and were not evident in the foot tissue, possibly because nonsense-mediated decay is obscured in actively transcribing tissues. Our work provides a set of control genes that may be useful to the wider community and illustrates how these genes may be used to assay differences in expression in a variety of tissues.

Citation

Johnson, H. F., & Davison, A. (2019). A new set of endogenous control genes for use in quantitative real-time PCR experiments show that formin Ldia2dex transcripts are enriched in the early embryo of the pond snail Lymnaea stagnalis (Panpulmonata). Journal of Molluscan Studies, 85(4), 388-396. https://doi.org/10.1093/mollus/eyz027

• Journal Article Type: Article
• Acceptance Date: Jan 28, 2019
• Online Publication Date: Oct 9, 2019
• Publication Date: Nov 4, 2019
• Deposit Date: Jun 7, 2020
• Publicly Available Date: Aug 4, 2020
• Journal: Journal of Molluscan Studies
• Print ISSN: 0260-1230
• Electronic ISSN: 1464-3766
• Publisher: Oxford University Press (OUP)
• Peer Reviewed: Peer Reviewed
• Volume: 85
• Issue: 4
• Pages: 388-396
• DOI: https://doi.org/10.1093/mollus/eyz027
• Keywords: Animal Science and Zoology; Aquatic Science
• Public URL: https://nottingham-repository.worktribe.com/output/4214937
• Publisher URL: https://academic.oup.com/mollus/article-abstract/85/4/389/5584423?redirectedFrom=fulltext