Reference Gene Selection for Real-Time Quantitative PCR Analysis of the Mouse Uterus in the Peri-Implantation Period
Open Access
- 24 April 2013
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 8 (4), e62462
- https://doi.org/10.1371/journal.pone.0062462
Abstract
The study of uterine gene expression patterns is valuable for understanding the biological and molecular mechanisms that occur during embryo implantation. Real-time quantitative RT-PCR (qRT-PCR) is an extremely sensitive technique that allows for the precise quantification of mRNA abundance; however, selecting stable reference genes suitable for the normalization of qRT-PCR data is required to avoid the misinterpretation of experimental results and erroneous analyses. This study employs several mouse models, including an early pregnancy, a pseudopregnancy, a delayed implantation and activation, an artificial decidualization and a hormonal treatment model; ten candidate reference genes (PPIA, RPLP0, HPRT1, GAPDH, ACTB, TBP, B2M, 18S, UBC and TUBA) that are found in uterine tissues were assessed for their suitability as internal controls for relative qRT-PCR quantification. GeNormPLUS, NormFinder, and BestKeeper were used to evaluate these candidate reference genes, and all of these methods identified RPLP0 and GAPDH as the most stable candidates and B2M and 18S as the least stable candidates. However, when the different models were analyzed separately, the reference genes exhibited some variation in their expression levels.This publication has 25 references indexed in Scilit:
- Molecular aspects of implantation failureBiochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2012
- Variation in stability of housekeeping genes in endometrium of healthy and polycystic ovarian syndrome womenHuman Reproduction, 2011
- Selection of reference genes for studies of porcine endometrial gene expression on gestational day 12Biochemical and Biophysical Research Communications, 2011
- A practical approach to RT-qPCR—Publishing data that conform to the MIQE guidelinesMethods, 2010
- How to do successful gene expression analysis using real-time PCRMethods, 2010
- Endometrial Decidualization: Of Mice and MenSeminars in Reproductive Medicine, 2010
- Identification of Suitable Normalizing Genes for Quantitative Real-Time RT-PCR Analysis of Gene Expression in Fetal Mouse GonadsSexual Development, 2009
- Sensitivity of housekeeping genes in the hypothalamus to mismatch in diets between pre- and postnatal periods in miceNeuroscience Letters, 2008
- Mouse models of implantationTrends in Endocrinology & Metabolism, 2007
- Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genesGenome Biology, 2002