Blastocyst transfer in mice alters the placental transcriptome and growth.

Assisted reproduction technologies (ART) are becoming increasingly common. Therefore, how these procedures influence gene regulation and feto-placental development are important to explore. Here, we assess the effects of blastocyst transfer on mouse placental growth and transcriptome. C57Bl/6 blastocysts were transferred into uteri of B6D2F1 pseudopregnant females and dissected at embryonic day 10.5 for analysis. Compared to non-transferred controls, placentas from transferred conceptuses weighed less even though the embryos were larger on average. This suggested a compensatory increase in placental efficiency. RNA-sequencing of whole male placentas revealed 543 differentially expressed genes (DEGs) after blastocyst transfer: 188 and 355 genes were down-regulated and up-regulated, respectively. DEGs were independently validated in male and female placentas. Bioinformatic analyses revealed that DEGs represented expression in all major placental cell types and included genes that are critical for placenta development and/or function. Furthermore, the direction of transcriptional change in response to blastocyst transfer implied an adaptive response to improve placental function to maintain fetal growth. Our analysis revealed that CpG methylation at regulatory regions of two DEGs was unchanged in female transferred placentas and that DEGs had fewer gene-associated CpG islands (within ~20 kb region) compared to the larger genome. These data suggested that altered methylation at proximal promoter regions might not lead to transcriptional disruption in transferred placentas. Genomic clustering of some DEGs warrants further investigation of long-range, cis-acting epigenetic mechanisms including histone modifications together with DNA methylation. We conclude that embryo transfer, a protocol required for ART, significantly impacts the placental transcriptome and growth.This work was supported by grants from the Centre for Trophoblast Research (CTR) (to EDW), Lister Institute for Preventative Medicine (to EDW), and Canadian Institutes for Health Research (to JCC). KM was funded by a Newnham College (Cambridge) studentship and A.G. Leventis scholarship. SJT was funded by a Next Generation Fellowship (CTR). GETB was funded by a Wellcome Trust PhD studentship in Developmental Mechanisms. RSH and MP were funded by the CTR