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Keiichiro Kizaki, Koichi Ushizawa, Miki Kimura, Kosuke Iga, Toru Takahashi, Kazuyoshi Hashizume, Active Roles of Heparin-Binding EGF-Like Growth Factor and Its Receptor in Bovine Endometrium During Implantation., Biology of Reproduction, Volume 83, Issue Suppl_1, 1 November 2010, Page 459, https://doi.org/10.1093/biolreprod/83.s1.459
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Heparin-binding EGF-like growth factor (HBEGF) participates in a wide range of physiological and pathological processes, including heart development, wound healing, tumor development and angiogenesis. HBEGF has also been highlighted as an early molecular marker of conceptus-endometrium cross-talk during implantation. Although expression and signaling system of HBEGF and its receptor (EGFR) have been identified in human and mice, there are still unclear in cattle. The aim of this study was to identify the expression of HBEGF and EGFR in bovine endometrium, and investigate the role of this system during implantation and placentation in cattle. The endometrial tissues were collected from Japanese Black cow from early to late gestation and estrous cycle. The gene expression of HBEGF and EGFR were measured by quantitative real-time RT-PCR (QPCR), and the localization was examined by in situ hybridization. In QPCR analysis, expression of HBEGF mRNA was detected in endometrium during the estrous cycle, and the abundance of mRNA increased in intercaruncular endometrium during gestation, but not in caruncular endometrium. On the other hand, EGFR mRNA also expressed in estrous cycle, and strongly increased in caruncle during gestation. In situ hybridization analysis showed that HBEGF mRNA was detected in endometrial epithelia and stroma during estrous cycle, and strongly increased in luminal and glandular epithelium at day 19 of gestation. EGFR was weakly detected in endometrial epithelia and stroma at estrus, and mainly observed in luminal epithelium at during early gestation. Further we examined the role of HBEGF-EGFR system in bovine endometrium using cultured endometrial epithelial and stromal cells. Both genes were expressed in cultured endometrial eplithelial and stromal cells. Exogenously added recombinant HBEGF (0.1-10 ng/mL) stimulated dose-dependently the proliferation of endometrial stromal cells in vitro. When the cells were cultured for 4 days on different culture condition; on collagen-coated dish, on collagen gel and in collagen gel, respectively, the expression of EGFR was not changed in any conditions, but HBEGF was markedly increased in collagen gel culture. These results demonstrate that HBEGF-EGFR signaling system presents in bovine endometrium, and exhibits proliferation potential to the endometrial cells, indicating that the system play crucial role in implantation and placental development in cattle. This research was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports Science and Technology, Japan.
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