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Effect of lactation on conceptus-maternal interactions at the initiation of implantation in cattle: Effects on the conceptus transcriptome

ABSTRACT: The hypothesis tested was that the uterine environment of lactating cows would affect conceptus gene expression. Approximately 65-75 days post-partum (dpp) the estrous cycles of non-lactating (dried off immediately post partum: n=12) and lactating (n=13) cows were synchronized and on Day 7 a high quality blastocyst derived from superovulated heifers was transferred. A control group of maiden heifers (n=8) were synchronized, inseminated to a standing heat and slaughtered on the same day as non-lactating and lactating recipients (Day 19; estrus=Day 0). The ipsilateral uterine horn was flushed with 10 ml PBS and the conceptus, when present, and uterine luminal fluid (ULF) snap frozen in liquid nitrogen prior to analysis. Gene expression analysis of the conceptus was performed by RNA sequencing analysis while amino acid (aa) composition of ULF was determined by High Performance Liquid Chromatography (HPLC). Eight differentially expressed genes (DEGs) were identified between conceptuses recovered from non-lactating cows versus heifers while 269 DEGs (100 up-regulated and 169 down-regulated) were identified between conceptuses recovered from lactating cows compared to heifers. The aa, alanine, glycine, serine and threonine, arginine, leucine and valine, were significantly lower in abundance in ULF recovered from heifers compared to both non-lactating or lactating cows. Glutamic acid, glutamine and lysine concentrations were lowest in heifers compared to both cow groups. This study demonstrates that exposure of a grade one embryo to a uterine environment that has been exposed to the metabolic stresses associated with lactation modifies the transcriptome of the conceptus and aa composition of the ULF.

ORGANISM(S): Bos taurus

PROVIDER: GSE85563 | GEO | 2017/08/12

SECONDARY ACCESSION(S): PRJNA338805

REPOSITORIES: GEO

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Project description:In cattle, maternal recognition of pregnancy occurs on Day 16 via secretion of interferon tau (IFNT) by the conceptus. The endometrium can distinguish between embryos with different developmental competencies. In eutherian mammals, X-chromosome inactivation (XCI) is required to ensure an equal transcriptional level of most X-linked genes for both male and female embryos in adult tissues, but this process is markedly different in cattle than mice. We examined how sexual dimorphism affected conceptus gene expression and amino acid composition as well as the endometrial transcriptome during the peri-implantation period of pregnancy. Of the 5132 genes were differently expressed on Day 19 in male compared to female conceptuses, 2.7% were located on the X-chromosome. Concentrations of specific amino acids were higher in the uterine luminal fluid with male compared to female conceptuses, while female conceptuses had higher expression of specific amino acid transporters (SLC6A19 and SLC1A35). Of note, the endometrial transcriptome was not different in cattle gestating a male or a female conceptus. These data support the hypothesis that, far from being a blastocyst specific phenomenon, XCI is incomplete before and during implantation in cattle. Despite differences in gene expression and amino acid utilization in male versus female conceptuses, the sex of the conceptus itself does not elicit a different response in the endometrium. Following a synchronized estrous cycle, all heifers observed in standing estrus (=Day 0, n=30) were inseminated with semen from a proven sire. All samples were recovered at slaughter on Day 19 following estrus corresponding to the initiation of implantation in cattle, flushed with 10 ml of PBS and the presence of a conceptus was observed under a stereo-microscope (n=24). Each conceptus was dissected into 4 pieces, 3 containing only trophectoderm cells and one containing the embryonic disc along with associated trophectoderm cells, and immediately snap-frozen in liquid nitrogen along with the corresponding intercaruncular endometrium from the uterine horn ipsilateral to the corpus luteum. DNA was extracted from each conceptus with phenol/chloroform treatment and finally re-suspended in 200 Î¼L of milliQ water. Two microliters of each sample were used to perform embryo sexing by PCR amplification of sex-specific polymorphic fragments in the amelogenin gene. N=5 samples of intercaruncular endometirum and the corresponding trophectoderm only sample were anaylsed for gene expression.

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Effect of lactation on conceptus-maternal interactions at the initiation of implantation in cattle: Effects on the conceptus transcriptome

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