Abstract
This study compared the gene expression of extraembryonic membranes (EEM) from in vitro produced (IVP) and in vivo (AI) derived pregnancies. A piece of conceptus (day 18) or chorioallantois (day 32) was used for DNA and RNA isolation and sex determination. Male and female ratios were analyzed by Chi-square. A total of three samples per sex and group (AI and IVP, days 18 and 32) were used for transcriptome analysis. Differentially expressed genes (DEGs) were determined using edgeR-robust. A false discovery rate (FDR) <0.05 was used for statistical significance. Sex ratio was similar on day 18 for AI and IVP groups. On day 32, the IVP group had a greater number of females than males (75 vs. 25%, P = 0.004). When comparing AI and IVP males vs. females, in both groups, genes upregulated in females on day 18 were related to placental function such as PAGs and TKDPs. On males on day 18, IFNT-related genes were upregulated. Comparing the techniques within sex, on day 18 female conceptuses, 50 genes were upregulated in IVP, and 21 in AI. IGF2, which is involved in placenta development, and APOA2, APOB, and APOE, involved in lipid metabolism, were upregulated in IVP conceptuses. On day 18, males had 15 upregulated genes in AI and 7 in IVP. On day 32, females had 21 upregulated genes in AI and 53 in IVP. Genes involved in lipid synthesis and metabolism were increased in the IVP group. Males on day 32 presented 899 DEGs, 564 upregulated in AI and 335 in IVP. Embryos from IVP had decreased expression of genes related to lipid and carbohydrate metabolism. Interestingly, pregnancy-associated glycoproteins (PAG) 7, 9, 10, and 19, were downregulated in IVP male. In conclusion, IVP-derived male embryos were more susceptible to alterations in gene expression and these effects extend to the peri-implantation period including genes associated with placental development and markers of placental function.
Highlights
It is generally accepted that embryos produced in vitro (IVP) differ in quality than in vivo-derived (IVD) embryos in terms of ultrastructure (Rizos et al, 2002a), gene expression profiles (Lonergan et al, 2003a,b; Gad et al, 2012), and cryotolerance (Rizos et al, 2002c)
When comparing the viability of in vitro produced (IVP) and IVD conceptuses on day 16 of pregnancy, more degenerated conceptuses (25 [7/28] vs. 5% [2/34]) and smaller embryonic discs were found in the IVP group compared to their IVD counterparts (Bertolini et al, 2002b)
PAG1 was found increased in the male conceptuses (Figure 3B), as well as genes related to maternal recognition of pregnancy, such as IFNAR1, IFNTAU, IFNT, and IFNT2
Summary
It is generally accepted that embryos produced in vitro (IVP) differ in quality than in vivo-derived (IVD) embryos in terms of ultrastructure (Rizos et al, 2002a), gene expression profiles (Lonergan et al, 2003a,b; Gad et al, 2012), and cryotolerance (Rizos et al, 2002c). When comparing the viability of IVP and IVD conceptuses on day 16 of pregnancy, more degenerated conceptuses (25 [7/28] vs 5% [2/34]) and smaller embryonic discs were found in the IVP group compared to their IVD counterparts (Bertolini et al, 2002b). They seem to induce different endometrial gene expression during the peri-implantation period (15–20 days after fertilization), altering expression of genes associated with immune response and metabolism (Bauersachs et al, 2009; Mansouri-Attia et al, 2009; Sandra et al, 2011; Mathew et al, 2019). The increased number of losses of IVP pregnancies between days 30 and 90 have been attributed to some extent to reduced development of placental membranes and placental blood vessels (Hill et al, 2000), in part due to abnormal epigenetic reprogramming caused by the IVP systems (Bertolini et al, 2002a,b)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
