Antigestagens Mediate the Expression of Decidualization Markers, Extracellular Matrix Factors and Connexin 43 in Decidualized Dog Uterine Stromal (DUS) Cells

SIMPLE SUMMARY: Adequate embryo-maternal communication is essential for a successful pregnancy. In the dog, this interaction is intimately associated with maternal stroma-derived decidual cells, the only cell population in the canine placenta expressing the nuclear progesterone receptor (PGR) and, therefore, sensitive to the circulating progesterone levels. Prepartum decrease of progesterone or clinical application of PGR blockers (antigestagens, e.g., aglepristone and mifepristone) induce placental release of luteolytic factors and terminate pregnancy. However, the importance of progesterone for decidual cells functionality has not been fully elucidated. Therefore, we investigated the effects of PGR blockers on the expression of markers of decidualization and cellular viability, as well as on epithelial and mesenchymal factors in in vitro decidualized dog uterine stromal (DUS) cells. Decidualization increased the expression of the respective markers, including factors involved in cell growth and prostaglandin synthesis. Their expression was suppressed by the application of antigestagens. Additionally, the expression of factors involved in tissue remodeling and cell-cell communication was increased, and antiproliferative and proapoptotic effects were induced in decidualized cells. Altogether, progesterone signaling appears to be crucial for modulating decidual cells physiology and biological activity, and thus for the maintenance of pregnancy. ABSTRACT: Feto-maternal communication in the dog involves the differentiation of stromal cells into decidual cells. As the only placental cells expressing the nuclear progesterone (P4) receptor (PGR), decidual cells play crucial roles in the maintenance and termination of pregnancy. Accordingly, to investigate possible PGR-mediated mechanisms in canine decidual cells, in vitro decidualized dog uterine stromal (DUS) cells were treated with functional PGR-blockers, mifepristone and aglepristone. Effects on decidualization markers, epithelial and mesenchymal factors, and markers of cellular viability were assessed. Decidualization increased the expression of PTGES, PGR, IGF1, and PRLR, along with ECM1, COL4 and CX43, but downregulated IGF2. DUS cells retained their mesenchymal character, and the expression of COL4 indicated the mesenchymal-epithelial transformation. Antigestagen treatment decreased the availability of PTGES, PRLR, IGF1 and PGR. Furthermore, antigestagens decreased the mRNA and protein expression of CX43, and transcriptional levels of ECM1 and COL4. Additionally, antigestagens increased levels of activated-CASP3 (a proapoptotic factor), associated with lowered levels of PCNA (a proliferation marker). These data reveal important aspects of the functional involvement of PGR in canine decidual cells, regarding the expression of decidualization markers and acquisition of epithelial-like characteristics. Some of these mechanisms may be crucial for the maintenance and/or termination of canine pregnancy.