Imaging of Vascular Development in Early Mouse Decidua and Its Association with Leukocytes and Trophoblasts

In species with endometrial decidualization and hemochorial placentation (humans, mice, and others), leukocytes localize to early implant sites and contribute to decidual angiogenesis, spiral arterial remodeling, and trophoblast invasion. Relationships between leukocytes, trophoblasts, and the decidual vasculature are not fully defined. Early C57BL/6J implant sites were analyzed by flow cytometry to define leukocyte subsets and by whole-mount immunohistochemistry to visualize relationships between leukocytes, decidual vessels, and trophoblasts. Ptprc(+) (CD45(+)) cells increased in decidua between Gestational Day (GD) 5.5 and GD 9.5. Uterine natural killer (uNK) cells that showed dynamic expression of Cd (CD) 69, an activating receptor, and Klrg1 (KLRG1), an inhibitory receptor, localized mesometrially and were the dominant CD45(+) cells between GD 5.5 and GD 7.5. At GD 8.5, immature monocytes that occurred throughout decidua exceeded uNK cells numerically and many leukocytes acquired irregular shapes, and leukocyte-leukocyte conjugates became frequent. Vessels were morphologically heterogeneous and regionally unique. Migrating trophoblasts were first observed at GD 6.5 and, at GD 9.5, breached endothelium, entered vascular lumens, and appeared to occlude some vessels, as described for human spiral arteries. No leukocyte-trophoblast conjugates were detected. Whole-mount staining gave unparalleled decidual vascular detail and cell-specific positional information. Its application across murine models of pregnancy disturbances should significantly advance our understanding of the maternal-fetal interface.