Abstract
BackgroundExtracellular matrix metalloproteinase inducer (EMMPRIN) regulates several biological functions involving the modulation of cell behaviors via cell-cell and cell-matrix interactions. According to its diverse functions, we hypothesized that EMMPRIN may play an important role in endometrial remodeling and establishment of pregnancy in cow.MethodsIn this study, endometrial tissues from the cyclic cows during before ovulation, after ovulation and middle of estrous cycle; and pregnant endometrial tissues from Day 19 to 35 of gestation have been used. Expression of mRNA was analyzed by RT-PCR, qPCR and in situ hybridization whereas protein expression by immunohistochemistry and western blot analysis.ResultsEMMPRIN mRNA was expressed in both cyclic and pregnant endometrium and significantly higher in the endometrium at Day 35 of gestation than the cyclic endometrium. In Western blot analysis, an approximately 65 kDa band was detected in the endometrium, and approximately 51 kDa in the cultured bovine epithelial cells and BT-1 cells, respectively. Both in situ hybridization and immunohistochemistry data showed that EMMPRIN was primarily expressed in luminal and glandular epithelium with strong staining on Day 19 conceptus. At Day 19 of gestation, expression of EMMPRIN mRNA on luminal epithelium was decreased than that observed at middle of estrous cycle, however, on Day 30 of gestation, slightly increased expression was found at the site of placentation. Expression of matrix metalloproteinase-2 (MMP-2) and MMP-14 mRNA were mainly detected in stroma and their expression also decreased at Day 19 of gestation however it was also expressed at the site of placentation at Day 30 of gestation as observed for EMMPRIN. Expression of MMP-1 or -9 mRNA was very low and was below the detection limit in the cyclic and pregnant endometrium.ConclusionEMMPRIN from the luminal epithelium may regulate the expression of stromal MMP-2 and -14 suggesting its crucial role in adhesion and fusion of embryo to luminal epithelium by directly itself through physiological tissues remodeling and developmental process, and/or stimulating MMPs to compensate endometrial functions.
Highlights
Extracellular matrix metalloproteinase inducer (EMMPRIN) regulates several biological functions involving the modulation of cell behaviors via cell-cell and cell-matrix interactions
Expression and localization of EMMPRIN mRNA in the bovine endometrium A partial cDNA for bovine EMMPRIN was amplified by RT-PCR from endometrial tissue after being isolated in the middle of the estrous cycle (Figure 1A)
The expression of EMMPRIN mRNA was analyzed by quantitative RT-PCR (qPCR) during the estrous cycle and at Day 35 of pregnancy (Figure 1B)
Summary
Extracellular matrix metalloproteinase inducer (EMMPRIN) regulates several biological functions involving the modulation of cell behaviors via cell-cell and cell-matrix interactions. The endometrium needs to be prepared for this event during the estrous cycle, and especially around implantation; and secondly, the trophoblast must be able to fuse with the endometrial epithelium in the caruncle region. A number of molecules are expressed at the embryo-maternal interface including interferon-tau, cytokines, growth factors, hormones, and MMP [8,9,10,11] These changes in the endometrium are partly modulated by the expression of the MMP system [12,13,14], a disintegrin and metalloproteinase with thrombospondin motif (ADAMTS)-1 [15,16], and extracellular matrix metalloproteinase inducer (EMMPRIN) [17] in coordination with ovarian steroids [18,19]
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