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R. Wordinger, J. Nile, and G. Stevens

Summary. Pregnant CD-1 mice were injected with diethylstilboestrol (10μg/kg body weight) in 0·1 ml maize oil, or maize oil alone, on Day 16 of gestation. Six experimental and 6 control female progeny were killed daily from birth until Day 7 and uterine tissues were examined by light microscopy. In-utero exposure to diethylstilboestrol resulted in hypertrophy of luminal epithelial cells and premature formation of uterine glands. The initial sign of uterine gland formation was invagination of the uterine surface epithelial cell layer into the underlying connective tissue stroma. A temporal difference occurred between control animals and those exposed to diethylstilboestrol: uterine gland formation first occurred in experimental progeny on Day 4, but not until Day 5 in control progeny. Uterine glands which extended deep into the connective tissue stroma to the myometrium were present in diethylstilboestrol-treated progeny by Day 7, but remained in the superficial endometrial connective tissue stroma in control animals. The results indicate that prenatal exposure of mice to diethylstilboestrol causes uterine epithelial cell hypertrophy at birth and the premature formation of uterine glands during the first week of neonatal uterine development.

Keywords: uterine glands; development; diethylstilboestrol; mouse

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R. J. Wordinger, J. F. Dickey, and A. R. Ellicott

Limited histochemical information is available concerning the lipid content of the bovine oviduct (Weeth & Herman, 1952; Bjorkman & Fredricsson, 1961) and endometrium (Weeth & Herman, 1952; Skjerven, 1956; Marinov & Lovell, 1968), and the proximity of the corpus luteum or developing follicle has not been examined as a possible local influence on such content. The object of this study was to examine the influence of the stage of the oestrous cycle as well as the proximity of the corpus luteum or developing follicle on the histochemically demonstrable lipid of the bovine oviduct and endometrium.

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R. J. Wordinger, A.-M. Brun-Zinkernagel, and T. Jackson

Summary. Guinea-pig (intrusive) and mouse (displacement) blastocysts display different cellular mechanisms of implantation. Blastocysts were placed in CM RL-1066 supplemented with either 10 or 20% fetal calf serum, 0·1m l-glutamine and antibiotics and then transferred to dishes previously coated with either Matrigel™ or type I collagen. After culture for 48 or 72 h, the dishes were processed for transmission electron microscopy. Blastocysts had attached to both extracellular matrices by 48 h. Matrigel™ elicited minimal trophoblast cell activity. Trophoblast cell projections were oriented parallel to the Matrigel™ and displayed little invasive activity, but trophoblast cells displayed active interaction with type I collagen. By 72 h, trophoblast cells exhibited slender, anastomosing projections which extended into the collagen matrix. Bundles of microfilaments running parallel with the long axis of the projections were observed. The morphology of type I collagen was altered in the immediate vicinity of the trophoblast projections. The projections interdigitated and desmosomes developed between processes. Projections appeared to meet, fuse and entrap matrix. These results suggest that trophoblast cells do not significantly interact with Matrigel™, but penetrate into type I collagen.

Keywords: trophoblast; extracellular matrices; ultrastructure; in vitro; guinea-pig; mouse

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R. J. Wordinger, D. Brown, E. Atkins, and F. L. Jackson

Summary. Pregnant mice were injected subcutaneously with diethylstilboestrol (DES: 10 μg/kg body weight in 0·1 ml corn oil) or corn oil alone on Day 15 or 16 of gestation (Day 1 = day of copulatory plug) and allowed to give birth. Female progeny from control and DES-exposed animals were superovulated with exogenous gonadotrophins at 6–8 weeks of age. In-vivo results indicated that the total number of ovulated ova, 2-cell embryos and blastocysts were significantly increased in DES-exposed progeny but that there was a decline in developmental potential from the ovulated ova stage to the blastocyst stage in these animals. However, there was no significant difference in the in-vitro development of 2-cell embryos to the blastocyst stage between control and DES-exposed animals. These results indicate that the ovaries of mice exposed in utero to DES are capable of responding to exogenous gonadotrophins and that second generation progeny have the potential for normal development to the early post-blastocyst stage of embryogenesis. The in-vivo decline in developmental potential may be attributable to reproductive tract abnormalities rather than ova/embryo defects.

Keywords: diethylstilboestrol; mouse; embryo; in vitro; development

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R. J. Wordinger, F. L. Jackson, and A. Morrill

Summary. The intraluminal injection of oil produced deciduoma formation in ovariectomized, mast cell-normal (+ /+) and mast cell-deficient (W/Wv) mice that were treated with exogenous steroids. Oil injection and trauma (e.g. sutures) also produced a deciduoma in ovariectomized + / + and W/Wv mice that had received a single control (+ / +) ovary transplanted under the kidney capsule. After transfers of donor blastocysts, implantation and live births were obtained in +/+ and W/Wv mice containing a single ovary transplant. Our results demonstate that uterine mast cells are not required for the production of a decidual cell response, implantation, gestation or the birth of live offspring in mice.

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R. J. Wordinger, E. L. Orr, K. Pace, L. Oakford, and A. Morrill

Summary. The ovaries from mast cell-normal (+/+) and mast cell-deficient (W/Wv) mice were examined with light and electron microscopy. In addition the effect of ovariectomy and subsequent steroid treatment on total uterine histamine content, total mast cell numbers and surface and glandular epithelial cell heights was measured. The ovaries of +/+ mice were normal, displaying various stages of follicular growth and atresia and numerous corpora lutea; the ovaries of W/Wv mice lacked follicles and corpora lutea but contained numerous hyperplastic interstitial cells which contained numerous lipid droplets, vesiculated mitochondria and abundant endoplasmic reticulum suggestive of steroid synthesis. Steroid treatment of ovariectomized +/+ and W/Wv mice caused a significant increase in uterine wet weight and endometrial surface and glandular epithelial cell heights. In +/+ mice, steroid treatment caused a concomitant increase in total mast cells per uterine horn while mast cells were totally absent in W/Wv mice. The increase in uterine histamine in +/+ mice is consistent with the increase in mast cell numbers. Measurable amounts of uterine histamine, which increases slightly after steroid treatment, were demonstrated in W/Wv mice. Since the uteri of +/+ and W/Wv mice respond to steroids in a similar manner with the sole exception being histamine content and mast cell numbers, our results demonstrate the potential of using these animals to investigate the role(s) of uterine mast cells and non-mast cell uterine histamine in the process of implantation and the formation of a decidual cell response.

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R. J. Wordinger, A. E. Moss, T. Lockard, D. Gray, I-F. C. Chang, and T. L. Jackson

Summary. Uterine samples were either rapidly frozen in liquid nitrogen or placed in Bouin's fixative. A commercial primary polyclonal antibody made in rabbits against human recombinant basic fibroblast growth factor (bFGF) was used. Western blot analysis indicated that the antibody was specific for bFGF and did not react with acidic FGF. The primary antibody was followed by either goat anti-rabbit immunoglobulin G (IgG) conjugated to the fluorescent phycobiliprotein tracer phycoerythrin or biotinylated goat anti-rabbit IgG and a biotin–avidin–peroxidase complex. Specificity controls using adjacent sections were carried out by (i) substituting normal rabbit sera for the primary antisera, (ii) omitting the primary antisera or (iii) extracting sections with NaCl (2 mol l−1) prior to the immunochemical procedures. No binding of the antibody was observed with any of the specificity control sections. The connective tissue stroma and the basal lamina associated with uterine glandular and surface epithelial layers were positive for bFGF. Localization was not observed within surface or glandular epithelial cells. The basal lamina and endothelial cells associated with blood vessels within the uterus and the smooth muscle cells of the myometrium were positive for bFGF. There were no differences in uterine localization patterns or intensity during the oestrous cycle or after ovariectomy and steroid hormone supplementation. These studies demonstrate the specific localization of bFGF within the mouse uterus.

Keywords: basic fibroblast growth factor; mouse