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First published online 19 November 2003
doi: 10.1242/jcs.00826

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Embryonic signals direct the formation of tight junctional permeability barrier in the decidualizing stroma during embryo implantation

¹ Department of Pediatrics, Division of Reproductive and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232-2678, USA
² Laboratory of Animal Reproduction, Graduate School of Agriculture Science, Tohoku University, Sendai, Japan

View larger version (87K): [in a new window]   Fig. 1. Northern blot hybridizations of occludin, claudin-1 and ZO-2 mRNAs in the periimplantation mouse uterus. The mRNA levels were detected in total RNA samples obtained from whole mouse uteri from days 1-8 of pregnancy. A 2.3 kb transcript for occludin, a 4.0 kb and a 1.5 kb transcript for claudin-1, and a 5.2 kb transcript for ZO-2 were detected in the uterus. The blot was hybridized with rpL7 to confirm integrity, equal loading, and blotting of RNA samples.

View larger version (150K): [in a new window]   Fig. 2. In situ hybridizations of occludin, claudin-1 and ZO-2 mRNAs in the periimplantation mouse uterus. Dark-field photomicrographs of representative uterine sections on days 4-6 of pregnancy are shown at 40x magnifications. Am, antimesometrial side; bl, blastocyst; em, embryo; ge, glandular epithelium; le, luminal epithelium; m, mesometrial side; myo, myometrium; pdz, primary decidual zone; s, stroma; sdz, secondary decidual zone. Scale bar: 400 µm.

View larger version (124K): [in a new window]   Fig. 3. Immunocytochemistry of occludin, claudin-1 and ZO-2 (A) and cytokeratin 8 (B) proteins in the periimplantation mouse uterus. Photomicrographs of representative uterine sections on days 4-6 of pregnancy are shown at 100x magnifications. TROMA-1 antibody recognizes cytokeratin-8 protein. Am, antimesometrial side; bl, blastocyst; em, embryo; ge, glandular epithelium; le, luminal epithelium; m, mesometrial side; myo. myometrium; pdz, primary decidual zone; s, stroma; sdz, secondary decidual zone. Scale bar: 400 µm.

View larger version (123K): [in a new window]   Fig. 4. Immunocytochemistry of occludin, claudin-1, ZO-1 and ZO-2 proteins at the implantation site induced by E2 in P4-primed delayed mice. Photomicrographs of representative uterine sections are shown at 40x. Am, antimesometrial side; bl, blastocyst; em, embryo; ge, glandular epithelium; le, luminal epithelium; m, mesometrial side; myo, myometrium; pdz, primary decidual zone; s, stroma; sdz, secondary decidual zone. Scale bar: 400 µm.

View larger version (79K): [in a new window]   Fig. 5. Localization of X-gal staining in sections of normal blastocysts or trophoblast vesicles formed in vitro from two-cell ROSA26 embryos. (a) Section of a control blastocyst transferred to the oviduct for sectioning, and (b) section of a trophoblast (Tr) vesicle transferred to the oviduct for sectioning. Note absence of inner cell mass (ICM) from the section of Tr vesicle. Scale bar: 50 µm.

View larger version (90K): [in a new window]   Fig. 6. Immunocytochemistry of ZO-1 protein in a day-6 deciduum induced by intact blastocysts or trophoblast vesicles and deciduoma induced by oil. Photomicrographs of representative uterine sections are shown at 100x magnifications. (a) A normal implantation site with a implanting blastocyst; (b) a decidual area induced by oil (deciduomata), and (c) a decidual area induced by trophoblastic vesicle (Tr). Am, anti-mesometrial side; em, embryo; le, luminal epithelium; m, mesometrial side; pdz, primary decidual zone; sdz, secondary decidual zone; Tr, trophectoderm. Scale bar: 50 µm.

View larger version (93K): [in a new window]   Fig. 7. Distribution of FITC-labeled bovine IgG in the embryo-induced deciduum and oil-induced deciduomata on day 6 of pregnancy. Fluorescence photomicrographs (100x) of a representative embryo-induced deciduum (a) and oil-induced deciduoma (b). Am, antimesometrial side; em, embryo; le, luminal epithelium; m, mesometrial side; pdz, primary decidual zone; sdz, secondary decidual zone. Scale bar: 100 µm.