Production of Hyaluronan by the Trophectoderm is a Prerequisite for Mouse Blastocyst Attachment
| Autor: | Michal Neeman, Ron Hadas, Nava Dekel, Michal Elbaz, Shifra Ben-Dor, Eran Gershon, Fortune Kohen, Aviad Cohen |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
biology urogenital system Chemistry Transgene Embryo Embryonic stem cell In vitro Cell biology 03 medical and health sciences 0302 clinical medicine medicine.anatomical_structure In vivo 030220 oncology & carcinogenesis embryonic structures medicine biology.protein Blastocyst Osteopontin Receptor reproductive and urinary physiology 030304 developmental biology |
| DOI: | 10.1101/2020.03.27.012880 |
| Popis: | Embryo implantation requires execution of highly synchronized processes at the feto-maternal interface, initiated by blastocyst attachment to the endometrial epithelium. Hyaluronan is a major ECM component known to regulate adhesion-associated biological processes in various physiological settings. We hypothesized that hyaluronan may facilitate blastocyst attachment. In order to test our hypothesis, we characterized the blastocyst expression of hyaluronan synthesizing and degrading enzymes, as well as the expression of hyaluronan receptors during attachment. The functional impact of hyaluronan was challenged by the use of mouse transgenic blastocysts, in which genes encoding for hyaluronan synthesizing enzymes were deleted using lentiviral incorporation of Cas-9 endonuclease alongside specific short-guide RNAs into the embryonic trophectoderm. Embryos with transgenic trophectoderm were tested for their attachmentin vitro, or assessed for implantationin vivo, upon transfer to foster dams. Deletion of the trophectoderm hyaluronan biosynthesis significantly reduced the number of blastocysts attached to human uterine epithelium cellsin vitro. Reduced attachment was also observedin vivo, in pregnant mice carrying blastocysts with hyaluronan-depleted trophectoderm. In agreement, trophectoderm expression of osteopontin, was downregulated upon depletion of hyaluronan. MRI measurements revealed a decrease in uterine blood vessels permeability. Uterine expression of VEGF-A, PTGS-2 and uterine osteopontin, which constitute the immediate response to blastocyst attachment was also reduced. Furthermore, impaired implantation, associated with a decrease in hyaluronan synthesis in the mural trophectoderm, obtained upon tamoxifen treatment, has been recovered by LIF administration. These results demonstrate that estrogen-regulated hyaluronan-synthesis in the trophectoderm is indispensable for mouse blastocysts attachment to the uterine epithelium. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|