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Single-cell atlas of the pregnant equine endometrium before and after implantation.

ABSTRACT: Embryo implantation in the mare occurs just over one month after fertilization, coinciding with the production of chorionic gonadotropin. The factors that regulate this late implantation in the mare, and whether they are unique or shared with more invasive embryos, remains poorly understood. This study aimed to determine and compare the transcriptome and subpopulations of endometrial cells before and after embryo implantation in the horse. Single-cell RNA sequencing was used to characterize the transcriptome of nearly 97 000 endometrial cells collected from biopsies of the endometrium at the beginning (day 33 of gestation) and the end of the embryo implantation (day 42 of gestation) in mares. Sixteen immune and 24 non-immune cell clusters were identified representing all known main cell populations, as well as subpopulations of horse immune cells such as resident innate lymphoid cells and mucosal-associated invariant T cells. Contrary to current knowledge, endometrial natural killer (eNK) cells were the most abundant endometrial leukocyte population during implantation in horses. Moreover, eNK cells not only expressed genes which may interact with fetal MHC I such as LY49F but also exert immunoregulatory functions regardless of MHC I expression such as CD96/TIGIT. Analogous to other species studied, upregulation of CXCR4 was found in several subpopulations of immune cells. Our results suggest that despite distinctive and later placentation compared with humans, horses share some key similarities in the mechanisms of embryo implantation.

ORGANISM(S): Equus caballus

PROVIDER: GSE279241 | GEO | 2024/10/15

REPOSITORIES: GEO

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Project description:The developmental origins of adult disease are now recognized to reflect intrauterine conditions during embryonic and fetal life. Cell-cell communication between the maternal endometrium and the pre-implantation embryo can occur by several means. Here, we show that maternal miRNAs are secreted by the endometrial epithelium to the endometrial fluid. Microarray assessments revealed the presence of specific miRNAs that are associated with the window of implantation and therefore in direct contact with the human preimplantation embryo. These miRNAS are transported as free or exosome-associated molecules secreted to the endometrial fluid and then uptake into the pre-implantation embryo through the trophoectoderm. Finally, these maternal miRNAs were able to induce transcriptional and functional modifications of the embryo. Therefore, we propose an innovative model whereby endometrial maternal miRNAS may function as transcriptomic regulators during early embryo development offering a new perspective on the developmental origins of adult diseases such as obesity, type 2 diabetes, and others that are now recognized to reflect intrauterine conditions. The B6C3F1 mice strain was purchased. Female mice aged 6M-bM-^@M-^S8 weeks were primed to ovulate by administering 10 IU pregnant mare serum gonadotropin(Sigma-Aldrich, Irvine, UK), followed by administration of 10 IU hCG(Sigma-Aldrich, Irvine, UK), 48 hours later. Females were housed overnight with male breeders and examined the following morning for the presence of a vaginal plug (day 1 of pregnancy). On day 2 of pregnancy, mice were sacrificed by cervical dislocation, and embryos flushed with PBS from the oviduct using a blunt 30-gauge needle. Then, embryos were cultured in CCM-30 medium (Vitrolife, Lubeck, Germany) in presence or absence of 400nM of mir-30d MIMIC or Scramble-Alexa 488 for 72 hours. Then embryos are four times washed in fresh CCM. Experiment were carried out by triplicate and with 30 embryos/condition

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Single-cell atlas of the pregnant equine endometrium before and after implantation.

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