Mary Bitner Anderson

Placental Endocrine Disruption Induced by Cadmium: Effects on P450 Cholesterol Side-Chain Cleavage and 3β-Hydroxysteroid Dehydrogenase Enzymes in Cultured Human Trophoblasts

Abstract We previously suggested that cadmium (Cd), an environmental toxicant and constituent of tobacco smoke, inhibits progesterone secretion in cultured human placental trophoblasts by inhibiting low-density lipoprotein receptor mRNA expression. In the current study, we investigated whether Cd also disrupts progesterone synthesis via P450 cholesterol side-chain cleavage (P450scc) and 3β-hydroxysteroid dehydrogenase (3β-HSD), enzymes that play important roles in placental steroidogenesis. Human cytotrophoblasts were purified by density gradient centrifugation and incubated in Dulbecco modified Eagle medium + 10% fetal bovine serum with 0, 5, 10, or 20 μM CdCl2 for 96 h. Cells progressed to syncytiotrophoblastic maturity regardless of treatment. No differences (P > 0.05) in cell protein and lactate dehydrogenase activity were observed between untreated trophoblasts and those treated with CdCl2. However, P450scc and 3β-HSD mRNA transcript levels declined in a dose-dependent manner (P <0.05) in trophoblasts cocultured with 5, 10, or 20 μM CdCl2. P450scc activity was similarly inhibited (P < 0.05) by CdCl2 treatment, although 3β-HSD activity was not significantly affected. Coculture with 8-bromo-cAMP enhanced progesterone secretion in untreated cultures but did not reverse the decline in progesterone secretion induced by CdCl2 treatment. CdCl2 failed to influence cAMP content in cultured cells. Collectively, results suggest that P450scc enzyme is another site at which Cd interferes with placental progesterone production. However, it is unlikely that an inhibition of cAMP is involved with the inhibition of progesterone biosynthesis by Cd in human trophoblasts.

Effects of acute and chronic exposure to cobalt on male reproduction in mice

Chronic exposure of male mice to cobaltous chloride dramatically affected their reproductive potential, while acute administration had minimal effects. Acute exposure, followed by evaluation weekly over a 7-week period, revealed no significant changes in epididymal sperm concentration or testicular weight. However, small but significant decreases in fertility at weeks 2 and 3 of the study were observed. Sperm motility was depressed only during the first week of the study. In chronic studies, cobalt affected fertility in a time- and dose-dependent manner. There was a decrease in testicular weight, epididymal sperm concentration, and fertility. Sperm motility was also depressed. Serum testosterone levels were dramatically increased in cobalt treated animals, while FSH and LH serum levels were normal. It appears that cobalt is directly or indirectly interfering with spermatogenesis and with local regulatory mechanisms in testosterone synthesis.

Multiple sites of action of the vitamin D endocrine system: FSH stimulation of testis 1,25-dihydroxyvitamin D3 receptors.

1,25-Dihydroxyvitamin D [1,25(OH)2D] receptors exist in numerous unexpected tissues. These include, for example, rat lung, heart, testis, and uterus, but not prostate and bladder. The issues of 1,25(OH)2D effects on and receptor location in the testis were addressed by (a) physiological and pharmacological manipulations of tubule cell types and (b) histological examination of testes of vitamin D-deficient rats. FSH treatment in hypophysectomized adult rats increased 1,25(OH)2D receptor levels by 135% (P less than 0.01). Busulfan treatment reduced testis receptor levels by 35% (P less than 0.05) after 35 days (maximum effect), and the effect was reversed after recovery (85 d). Cryptorchidism for 5 or 50 days resulted in modest (33%, P less than 0.05) or substantial (79%, P less than 0.001) reductions in receptor levels. Only the FSH treatment and 50 days cryptorchidism reduced receptor levels in the residual tissue. The testes of vit. D-deficient rats showed incomplete spermatogenesis and degenerative changes. Although interpretation is complicated by the intricate communication among testis cell types, these data suggest that the Sertoli cell is a primary site of action of 1,25(OH)2D in the testis. Moreover, these data indicate that 1,25(OH)2D receptor function in the testis relates to germ cell division/maturation, although this may be an indirect effect via the Sertoli cells.

Bioaccumulation of lead nitrate in red swamp crayfish (Procambarus clarkii)

Abstract Crayfish were exposed to intermediate concentrations of lead nitrate (150 μgl −1 and 1100 μgl −1 ) for periods up to 7 weeks. Lead clearance was monitored at 3 weeks following the 7 week exposure to the lower concentration. Lead bioaccumulation was demonstrated to be a time- and dose-dependent phenomenon in gills, hepatopancreas and abdominal muscle, but not the exoskeleton. The tissue concentrations of lead in soft tissues, in decreasing order were gills > hepatopancreas > muscle > hemolymph. Lead clearance was significant in all tissues evaluated except the hepatopancreas, the organ of metal storage and detoxification.

Bioaccumulation of chromium in red swamp crayfish (Procambarus clarkii)

Abstract Crayfish were exposed to a range of potassium dichromate concentrations (0.15, 0.30, 3.0 and 30 mg l−1) for periods up to 7 weeks. Chromium bioaccumulation in all tissues over the 7 week exposure period was not consistently time- and dose-dependent. The order of distribution of chromium into the various tissues was dependent upon the exposure concentration of the metal. Chromium clearance studies conducted 1 and 3 weeks following exposure demonstrated a concentration reduction in most tissues only at the highest exposure concentration of chromium (30 mg l−1). Histological studies demonstrated damage to both the gills and hepatopancreas at the lowest exposure concentration. The results suggest that the red swamp crayfish, Procambarus clarkii, is a useful biomarker for chromium exposure.

Protective action of zinc against cobalt-induced testicular damage in the mouse

The purpose of this study was to determine if toxic effects of cobalt on the murine testis could be prevented by zinc, an essential metal for spermatogenesis. CD-1 male mice were administered one of the following in their drinking water: 1) 400 ppm CoCl2, 2) 800 ppm ZnCl2, 3) 400 ppm CoCl2 + 800 ppm ZnCl2, or 4) distilled water. After 13 weeks, animals were sacrificed and testes were excised, weighed, and processed for histologic study. Comparison of testicular weights revealed no difference between the control and zinc-treated groups, while there was a small but significant reduction in the zinc/cobalt-treated group, and a large reduction in the cobalt-treated group. Histologic evaluation of testes confirmed the degenerative effects of cobalt, as well as the normal morphology in the zinc-treated group. Furthermore, 90% of the animals in the zinc/cobalt-treated group exhibited complete or partial protection as demonstrated by tubular morphology. This study indicates that zinc prevents cobalt-induced testicular damage.

Effects of cadmium on cell viability, trophoblastic development, and expression of low density lipoprotein receptor transcripts in cultured human placental cells

Previously, we have demonstrated that cadmium inhibits progesterone release in cultured human trophoblast cells. In the present study, we investigated potential mechanism(s) by which cadmium may elicit this effect. Cytotrophoblasts were obtained via enzymatic dispersion, purified by density gradient centrifugation, and cultured with increasing concentrations of cadmium. Cadmium-induced suppression of progesterone release seemed to be independent of cell death, as no significant decline in viability was observed with cadmium treatment. Further, immunocytochemical localization of cellular boundaries and nuclei indicated approximately 94% syncytial maturity was attained by both untreated and cadmium-treated cells, demonstrating that cadmium did not inhibit syncytial development. However, the abundance of LDL receptor (LDL-R) mRNA transcripts, as determined by competitive RT-PCR, was reduced (P < 0.05) by cadmium exposure in an apparent dose-dependent manner. Thus, the LDL-R, by which cholesterol substrate is supplied to the syncytiotrophoblast, is one site at which cadmium may interfere with placental progesterone production.

Cadmium accumulation and effects on progesterone release by cultured human trophoblast cells.

This study was designed to examine the characteristics of cadmium bioaccumulation by human trophoblast cells in culture and the subsequent effect of cadmium exposure on progesterone production and syncytial formation. The accumulation of cadmium suggested a time- and dose-dependent relationship, although it was not significant. The rate of metal accumulation was similar in all cadmium-treated groups. After 72 h of continuous exposure to cadmium concentrations of 5, 10, and 20 microM, progesterone release was diminished to 69, 51, and 38% of control values (P < 0.05), respectively. When cells were exposed to cadmium from 72 to 96 h (after syncytial development), progesterone release exhibited the same pattern of decline in response to increasing cadmium concentrations. Histologic evaluation of whole mounts of trophoblast cells exposed to 20 microM CdCl2 for 96 h revealed that syncytial formation seemed to be uninhibited. The pattern of cadmium-accumulation by normal cultured human trophoblast cells suggests a time- and dose-relationship with a concomitant decrease in progesterone release that occurs without apparent inhibition of syncytial development.

Murine strain differences and the effects of zinc on cadmium concentrations in tissues after acute cadmium exposure

Abstract The role of strain differences in cadmium tissue distribution was studied using sensitive (129/J) and resistant (A/J) mice. These murine strains have previously been shown to differ in their susceptibility to cadmium-induced testicular toxicity. Cadmium concentration was measured in testis, epididymis, seminal vesicle, liver, and kidney at 24 h after cadmium chloride exposure (4, 10, and 20 μmol/kg CdCl2). The 129/J mice exhibited a significant increase in cadmium concentration in testis, epididymis, and seminal vesicle at all cadmium doses used, compared to A/J mice. However, cadmium concentrations in liver and kidney were not different between the strains, at any dose, indicating that cadmium uptake is similar in these organs at 24 h. These murine strains demonstrate similar hepatic and renal cadmium uptake but significantly different cadmium accumulation in the reproductive organs at 24 h. The mechanism of the protective effect of zinc on cadmium toxicity was studied by assessing the impact of zinc acetate (ZnAc) treatment on cadmium concentrations in 129/J mice after 24 h. Zinc pretreatment (250 μmol/kg ZnAc), given 24 h prior to 20 μmol/kg CdCl2 administration, significantly decreased the amount of cadmium in the testis, epididymis, and seminal vesicle of 129/J mice, and significantly increased the cadmium content of the liver after 24 h. Cadmium levels in the kidney were unaffected at this time. Zinc pretreatment also prevented the cadmium-induced decrease in testicular sperm concentration and epididymal sperm motility seen in 129/J mice. These findings suggest that the differences in the two murine strains may be attributed partly to the differential accumulation of cadmium in murine gonads. This may be caused by strain differences in the specificity of cadmium transport mechanisms. The protective role of zinc in cadmium-induced testicular toxicity in the sensitive strain may be due to an interference in the cadmium uptake by susceptible reproductive organs.

Solubility and bioavailability of lead following oral ingestion of vitrified slagged aggregate

Abstract The objective of this study was to evaluate the solubility of lead from various lead salts and vitrified slagged aggregate (VSA) in aqueous solutions.