Abstract
As sport career is a priority in most of equine breeds, mares are frequently bred for the first time at an advanced age. Both age and first gestation were shown to have a deleterious effect on reproduction outcomes, respectively on fertility and offspring weight but the effect mare’s parity in older mares on embryo quality has never been considered. The aim of this project was to determine the effect of old mare’s nulliparity on gene expression in embryos. Day 8 post ovulation embryos were collected from old (10-16 years old) nulliparous (ON, N=5) or multiparous (OM, N=6) non-nursing Saddlebred mares, inseminated with the semen of one stallion. Pure (TE_part) or inner cell mass enriched (ICMandTE) trophoblast were obtained by embryo bisection and paired end, non-oriented RNA sequencing (Illumina, NextSeq500) was performed on each hemi-embryo. To discriminate gene expression in the ICM from that in the TE, deconvolution (DeMixT R package) was used on the ICMandTE dataset. Differential expression was analyzed (DESeq2) with embryo sex and diameter as cofactors using a false discovery rate <0.05 cutoff. Although the expression of only a few genes was altered by mare’s nulliparity (33 in ICM and 23 in TE), those genes were related to nutrient exchanges and responses to environment signaling, both in ICM and TE, suggesting that the developing environment from these mares are not optimal for embryo growth. In conclusion, being nulliparous and old does not seem to be the perfect match for embryonic development in mares.
Summary sentence Mare’s parity in old mares impacts the expression of genes related to development and molecule exchanges in ICM and TE of blastocysts suggesting an adaptation to an altered environment.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
Grant support: This work was supported by the “Institut Français du Cheval et de l’Equitation” (grant numbers CS_2018_23, 2018). The National Research Institute for Agriculture, Food and Environment (INRAE) department Animal Physiology and Breeding Systems also supported this research.
https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE188866
- List of abbreviations
- DEG
- differential expressed genes
- DeMixT_ICM_cells
- deconvoluted gene expression in ICM cells
- DeMixT_TE_cells
- deconvoluted gene expression in TE cells
- ECM
- Extracellular matrix
- ERR
- embryo collection rate
- FDR
- false discovery rate
- GO BP
- Gene Ontology biological process
- GO
- Gene Ontology
- GSEA
- gene set enrichment analyses ICM: inner cell mass
- ICMandTE
- inner cell mass enriched hemi-embryo ICSI: intracytoplasmic sperm injection
- KEGG
- Kyoto Encyclopedia of Genes and Genomes Log2FC: log2 fold change
- NES
- normalized enrichment score
- OM
- old multiparous mares
- ON
- old nulliparous mares
- TE
- trophoblast
- TE_part
- pure trophoblast hemi-embryo
- XIST
- X inactive Specific Transcript