Functional Genetics and Development Division, The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Midlothian, UK.
Reprod Domest Anim. 2020 Jul;55(7):785-794. doi: 10.1111/rda.13685. Epub 2020 May 12.
Recent studies suggest associations exist between foetal size and sex, and gene expression at the porcine feto-maternal interface. It is essential to identify reference genes which have stable expression throughout gestation in feto-placental units associated with foetuses of different size and sex. qPCR was performed for 11 genes within porcine placentae and endometria at gestational days (GD) 30, 60 and 90. Several reference genes were found to have stable expression in these samples. The combination of B2m1 and Tbp1, and Hprt1 and Tbp1 had the most stable expression in endometria and placentae, respectively. Reference genes identified as having stable expression were utilized in a larger experiment with placentae and endometria associated with foetuses of different size and sex at four GD. The average expression of B2m1 and Tbp1 mRNAs was suitable for the normalization of temporal changes in endometria, and comparison between endometria supplying foetuses of different size throughout gestation. The average expression of Hprt1 and Tbp1 mRNAs was suitable for the normalization of placental mRNA expression for comparison of temporal changes and sex differences between placentae supplying foetuses of different sex throughout gestation. This combination was suitable for the normalization of mRNA expression in placentas supplying GD30, GD60 and GD90 foetuses of different size. This study has identified reference genes with stable expression in placentae and endometria across multiple gestational days, in tissues associated with foetuses of different size and sex. The results of these experiments highlight the importance of selecting appropriate reference genes for the biological comparison under investigation.
最近的研究表明，胎儿大小和性别与猪胎儿-母体界面的基因表达之间存在关联。在与不同大小和性别的胎儿相关的胎盘中鉴定整个妊娠期表达稳定的参考基因至关重要。在妊娠第 30、60 和 90 天，对猪胎盘和子宫内膜中的 11 个基因进行了 qPCR。在这些样本中发现了几个表达稳定的参考基因。B2m1 和 Tbp1 以及 Hprt1 和 Tbp1 的组合在子宫内膜和胎盘中的表达最稳定。鉴定为表达稳定的参考基因用于在更大的实验中使用，该实验涉及与不同大小和性别的胎儿相关的胎盘和子宫内膜，在四个 GD 中进行。B2m1 和 Tbp1 mRNA 的平均表达适合于子宫内膜中时间变化的归一化，以及整个妊娠期不同大小胎儿的子宫内膜之间的比较。Hprt1 和 Tbp1 mRNA 的平均表达适合于胎盘 mRNA 表达的归一化，用于比较整个妊娠期不同性别胎儿的胎盘之间的时间变化和性别差异。这种组合适合于不同大小的 GD30、GD60 和 GD90 胎儿的胎盘中 mRNA 表达的归一化。本研究鉴定了在多个妊娠天数、与不同大小和性别的胎儿相关的组织中表达稳定的胎盘和子宫内膜参考基因。这些实验的结果强调了为研究中的生物学比较选择合适参考基因的重要性。
