Environmental pollutants as aetiological agents in female reproductive pathology: placental glycan expression in normal and polychlorinated biphenyl (PCB)-exposed mink (Mustela vison).

Polychlorinated biphenyls (PCB) may cause growth retardation or fetal death in mink. Pathological changes in endotheliochorial mink placentae were examined following exposure to PCB during gestation. Placentae from six animals with average fetal crown-rump (C-R) lengths between 16 and 53 mm given 0.65 mg/day Clophen A50 (low dose), and one from five animals with an average fetal C-R length of 14 mm given 1.3 mg/day (high dose), were examined. Mink were treated from 9 to 24 days before mating until killed at day 53. Placentae were formalin-fixed with four size-matched controls and embedded in resin. Sections were stained with five biotinylated lectins to detect specific glycans. Both control and treated (low dose) mink showed degenerative changes in maternal endothelium from 13-16 mm, revealed by increased lectin binding, caused probably by high cell turnover during tissue remodelling. Controls of 47 and 50 mm exhibited fewer degenerate maternal endothelial cells. The 31-mm PCB-treated tissue showed separation of the trophoblast from the interstitial layer and, at 53 mm, loss of its normal architecture, increased damage to maternal endothelium and infarction. High-dose PCB was extremely toxic, producing fetal death or extensive placental infarction by 14 mm C-R length. Lectin staining thus revealed the effects of PCB toxicity, shown by increased injury to maternal endothelium and severe trophoblastic damage.

Histopathology of postpartum placental sites in mink (Mustela vison) exposed to polychlorinated biphenyls or fractions thereof.

Polychlorinated biphenyls (PCB) cause reproductive failure in mink. Ovulation and nidation occur, but the fetuses die during gestation. The toxicity of different chlorinated biphenyl (CB) congeners differs markedly. Dioxin-like congeners with no (0-ortho CBs) chlorine in the ortho position to the biphenyl bond are considered to be highly toxic. Altogether, 13 groups of 10 female mink (Mustela vison) were exposed to PCB or CB fractions thereof during the reproductive season of 1988 and 1989. In 1988, one group of mink received 2 mg/day of Clophen A50 and five groups received single fractions thereof or synthetic 0-ortho CB in their diet. In 1989, one group received 1.64 mg/day of Aroclor 1254 and six groups received combinations of fractions thereof. The daily amounts of the fractions administered per animal were equivalent to those present in 2 mg of Clophen A50 or 1.64 mg of Aroclor 1254. After administration for 3 months in both experiments, the animals were killed 5 days after parturition. Histological examination focused on the placental sites. The most involuted placental sites were those of early fetal death in primiparous, non-whelping animals in the 1989 experiment. The least involuted placental sites, displaying a hyperplastic and pleomorphic uterine luminal epithelium, in which proliferating cell nuclear antigen (PCNA) was present, were those of late fetal death in biparous, non-whelping animals in the 1988 experiment. The survival of the fetoplacental unit was related to PCB exposure. The histology of the placental sites seemed only to be related to the survival time of the fetuses and to the number of former reproductive seasons.

Polychlorinated biphenyl (PCB) exposure produces placental vascular and trophoblastic lesions in the mink (Mustela vison): a light and electron microscopic study.

Polychlorinated biphenyls (PCBs) cross the placenta and cause fetal death in mink. No indications of impaired implantation have been reported. To study the effects of PCB on mink placental morphology, 2 groups each of 10 animals were orally exposed to Clophen A50 at 0.65 mg (low dose) and 1.3 mg (high dose) per day for 54 days, starting before mating, with 10 control animals. Placentae from mid to late gestation were examined by light and electron microscopy. In the controls, 11% of placentae were degenerate compared to 31% (low dose) and 64% (high dose) in PCB-exposed mink. All control animals exhibited implantation sites, while one animal in the low dose and four in the high-dose group exhibited none. However, there was no difference between PCB-exposed and control animals in the number of placentation sites in implanted animals. Fetal death was markedly increased in PCB-exposed mink, with only four animals (low dose) having all viable fetuses and eight (low and high dose) having a mixture of viable and dead fetuses. Nine exposed animals displayed maternal vascular lesions in the placental labyrinthine zones of viable fetuses, comprising loss and degeneration of endothelial cells, thrombi and haemorrhages. Extracellular fluid was present between the interstitial layer of maternal vessels and the syncytiotrophoblast, and there was focal degeneration of the trophoblast and fetal vasculature. It appears, therefore, that exposure of the mink placenta to PCBs affects maternal vasculature and produces degenerative changes in the trophoblast and fetal vessels, leading to fetal growth retardation or death.

Histology of ovaries and uteri and levels of plasma progesterone, oestradiol-17beta and oestrone sulphate during the implantation period in mated and gonadotrophin-releasing hormone-treated mink (Mustela vison) exposed to polychlorinated biphenyls.

Earlier studies have shown that polychlorinated biphenyls (PCB) do not prevent ovulation, fertilization and implantation, but exposure of female mink during gestation caused fetal death. To understand this phenomenon, 30 PCB-exposed female mink and 30 controls were mated or induced to ovulate without fertilization by treatment with gonadotrophin-releasing hormone (GnRH) and correlations were measured between reproductive, morphological and endocrine parameters. The exposure to PCB (Aroclor 1254) started before ovulation. Equal numbers of animals from each group were killed on days 10, 17 and 26 post-coitum or on GnRH-injection days. Plasma concentrations of progesterone, oestradiol-17beta and oestrone sulphate in sampled blood were then measured. Ovaries, uteri and placentae were examined histologically. Compared with controls, exposure to PCB during the reproductive season resulted in significantly smaller uterine glandular diameters before implantation or at the end of the experiment in both mated and GnRH-treated mink. The GnRH treatment increased the sizes of the ovarian corpora lutea and oesterone sulphate 10 days after treatment but the increase in uterine glandular diameter was significant only in GnRH-treated control animals. Both GnRH-treated and mated PCB-exposed groups displayed a peak in oesterone sulphate concentration that was not observed in any of the control groups. Possible actions of PCB are discussed. The observed histological and hormonal changes in the mated PCB-exposed group did not prevent implantation. Exposure to PCB increased fetal mortality. This effect was associated with an effect on placental development.

Histology of uterine leiomyoma and occurrence in relation to reproductive activity in the Baltic gray seal (Halichoerus grypus).

A high prevalence of uterine leiomyoma has been reported in Baltic gray seals aged 15 years and above. Studies on Baltic seals during the 1970s revealed high tissue concentrations of the organochlorines bis(chlorophenyl)-1,1,1-trichloroethane (DDT) and polychlorinated biphenyls (PCBs), lowered reproduction rate, and pathologic changes. In the second half of the 1970s, decreases of PCB and DDT in Baltic biota occurred, and the prevalence of pregnancies in Baltic seals increased. Between 1975 and 1997, 53 Baltic gray seal females of age 15-40 years were found dead and sent to the Swedish Museum of Natural History. Seals were autopsied and 34/53 (64%) had uterine leiomyomas. Samples from 15 were sufficiently well preserved for histologic examination. Uterine leiomyomas were found most commonly in the uterine corpus but also were observed in the uterine horns, cervix, and vagina. Cut surfaces of the leiomyomas appeared as whorled white fibrous tissue. Histologically, spindle cells were arranged in a whorl-like pattern. The nuclei were rod-like and strikingly uniform in shape and size. Mitotic figures were rare. Immunohistochemical staining of the tumors showed a positive reaction to antibodies recognizing smooth muscle actin. Reproductively active gray seals have an ovarian corpus luteum or albicans for most of the year. In 22/34 (65%) gray seals with uterine leiomyomas, ovaries did not contain corpora. In gray seals without macroscopically detected uterine leiomyoma, ovaries from 6/19 (32%) seals had no corpora. It is possible that the development of leiomyoma in the seals is associated with organochlorines and the previous low reproductive activity.

Expression of the insulin-like growth factor II gene in polychlorinated biphenyl exposed female mink (Mustela vison) and their fetuses.

AIM: To study how polychlorinated biphenyls (PCBs) affect fetal growth and the expression of the insulin-like growth factor (IGF II) gene in the mink (Mustela vision). METHODS: Ten female mink were each exposed to 0.65 or 1.3 mg Clophen A50/day, respectively, during the reproductive season. The numbers and sizes of viable fetuses were recorded. The expression of the IGF II gene was studied by northern blotting using a mink specific IGF II cDNA probe. RESULTS: The number of viable fetuses decreased after PCB exposure in a dose dependent fashion. Expression of the IGF II gene in adult livers from PCB exposed animals was decreased, compared with control animals, in a dose dependent fashion. In contrast, IGF II expression in placentas and fetuses was unaltered. Furthermore, the maternal excretion of urinary cortisol increased in both exposed groups during the implantation period. CONCLUSIONS: Expression of the IGF II gene is downregulated by PCB exposure in the adult liver. There is also an indication that the regulation of the expression of this gene differs between adult and fetal life.

Insulin-like growth factor II in the mink (Mustela vison): determination of a cDNA nucleotide sequence and developmental regulation of its expression.

Mink cDNA for insulin-like growth factor II (IGF-II) has been isolated by cDNA synthesis from bulk mRNA and subsequent PCR-screening. Positive clones were sequenced and analyzed. Analysis of a cDNA revealed that, compared with human, the mature mink IGF-II peptide contains 68 instead of 67 amino acids with a serine insertion at residue 40. With this exception, the homology between the human and mink mature peptide is 100% and is 94% between rat and mink. Comparing IGF-II transcripts in fetal and adult mink liver, fetal tissue contains three transcripts of 5.8, 4.5, and 4.0 kb. In contrast, adult liver expresses low levels of a 4.6-kb transcript.