Validation of novel reference genes for RT‐qPCR studies of gene expression in Xenopus tropicalis during embryonic and post‐embryonic development. Issue 6 (29th April 2013)

• Record Type: Journal Article
• Authors: Dhorne‐Pollet, Sophie; Thélie, Aurore; Pollet, Nicolas
• Is Part Of: Developmental dynamics. Volume 242:Issue 6(2013:Jun.)
• Page Start: 709
• Page End: 717
• Publication Date: 2013-04-29
• Subjects: Morphogenesis -- Periodicals; Anatomy -- Periodicals; Anatomie -- Périodiques; Biologie du développement -- Périodiques; 571.833
• DOI: 10.1002/dvdy.23972
• Languages: English
• ISSNs: 1058-8388

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p> <underline>Background:</underline> Accurate interpretation of transcriptome profiling by quantitative PCR requires the establishment of species‐specific standards. However, the selection of reference genes for assessing RNA expression profiles in <italic>Xenopus laevis</italic> and <italic>Xenopus tropicalis</italic> was mostly based on historical reasons and they often only reflect the traditions of a laboratory. <underline>Results:</underline> We investigated the expression stability of 10 genes (<italic>dicer1, drosha, eef1a1, elavl3, gsc, h4, odc1, rpl8, smn2, tbp</italic>), 8 of which are commonly used as internal controls in published RT‐qPCR experiments. We defined specific primer pairs and evaluated their suitability as reference genes by performing RT‐qPCR expression profiling in <italic>Xenopus tropicalis</italic>. Gene expression stability was assayed in a set of 15 developmental stages from the egg to the froglet, and in dissected embryos. <underline>Conclusions:</underline> Overall, we determined a set of qualified reference genes for distinct developmental periods. We recommend the use of <italic>dicer1, drosha, eef1a1</italic>, and <italic>smn2</italic> from early embryonic development up to the end of metamorphosis. During early embryogenesis <italic>drosha, eef1a1, smn2</italic> are suitable. For the whole post‐embryonic development and for metamorphic stages including pro‐metamorphosis and metamorphic climax, we recommend the use of <italic>drosha</italic> and <italic>smn2</italic>. These reference genes should prove their usefulness for data comparison across studies. <italic>Developmental Dynamics 242:709–717, 2013</italic>. © 2013 Wiley Periodicals, Inc.</p> </abstract>