Abstract
During the preimplantation period of pregnancy in eutherian mammals, transcriptional and proteomic changes in the uterine endometrium are required to facilitate receptivity to an implanting blastocyst. These changes are mediated, in part, by proteins produced by the developing conceptus (inner cell mass and extraembryonic membranes). We hypothesized that this common process in early pregnancy in eutheria may be facilitated by highly conserved conceptus-derived proteins such as macrophage capping protein (CAPG). We propose that CAPG may share functionality in modifying the transcriptome of the endometrial epithelial cells to facilitate receptivity to implantation in species with different implantation strategies. A recombinant bovine form of CAPG (91% sequence identity between bovine and human) was produced and bovine endometrial epithelial (bEECs) and stromal (bESCs) and human endometrial epithelial cells (hEECs) were cultured for 24hours with and without recombinant bovine CAPG (rbCAPG). RNA sequencing and quantitative real-time PCR analysis were used to assess the transcriptional response to rbCAPG (Control, vehicle, CAPG 10, 100, 1000ng/mL: n=3 biological replicates per treatment per species). Treatment of bEECs with CAPG resulted in alterations in the abundance of 1052 transcripts (629 increased and 423 decreased) compared to vehicle controls. Treatment of hEECs with bovine CAPG increased expression of transcripts previously known to interact with CAPG in different systems (CAPZB, CAPZA2, ADD1, and ADK) compared with vehicle controls (P<.05). In conclusion, we have demonstrated that CAPG, a highly conserved protein in eutherian mammals, elicits a transcriptional response in the endometrial epithelium in species with different implantation strategies that may contribute to pregnancy success.
Highlights
In the majority of mammals, most pregnancy loss occurs in the pre- and peri-implantation periods of pregnancy
Individual tracts were sprayed with 70% of EtOH and the ipsilateral uterine horn (the horn attached to the ovary with the corpus luteum (CL)) was opened longitudinally to expose the uterine endometrium
We tested the hypothesis that capping protein (CAPG) is highly conserved among placental mammals and facilitates establishment of uterine receptivity and enhances pregnancy recognition in species with different implantation strategies, human and cow
Summary
In the majority of mammals, most pregnancy loss occurs in the pre- and peri-implantation periods of pregnancy. These heterogeneous cells respond spatially and temporally to steroid hormones, the concentrations of which fluctuate throughout the menstrual/estrous cycle in maternal circulation.[3,4] Following successful fertilization, the embryo enters the uterus on approximately day 4 of pregnancy where it is entirely reliant for its growth and survival on the secretion and transport of molecules from the luminal and glandular epithelial cells of the endometrium These molecules include growth factors, cytokines, amino acids and ions, and are collectively termed uterine luminal fluid (ULF).[5,6] ULF supports growth and development of the conceptus until a fully functional placenta is established.[7] The embryo undergoes a number of key morphological changes in the uterus prior to implantation including transition from a morula to a blastocyst, at which stage differentiation begins between the inner cell mass (that forms the fetus) and the outer cell mass (the trophectoderm, that goes on to form the placenta). Our specific aims were to (1) assess levels of sequence conservation of CAPG in representative eutherian mammal species, (2) produce a recombinant form of bovine CAPG, and (3) test how the cells of the endometrium from different species respond to this recombinant bovine CAPG (rbCAPG) in vitro
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