Microarray analysis of changes in bone cell gene expression early after cadmium gavage in mice

We developed an in vivo model for cadmium-induced bone loss in which mice excrete bone mineral in feces beginning 8 h after cadmium gavage. Female mice of three strains [CF1, MTN (metallothionein-wild-type), and MT1,2KO (MT1,2-deficient)] were placed on a low-calcium diet for 2 weeks. Each mouse was gavaged with 200 microg Cd or vehicle only. Fecal calcium was monitored daily for 9 days, beginning 4 days before cadmium gavage, to document the bone response. For CF1 mice, bones were taken from four groups: +/- Cd, 2 h after Cd and +/- Cd, 4 h after Cd. MTN and MT1,2KO strains had two groups each: +/-Cd, 4 h after Cd. PolyA+ RNA preparations from marrow-free shafts of femura and tibiae of each +/- Cd pair were submitted to Incyte Genomics for microarray analysis. Fecal Ca results showed that bone calcium excreted after cadmium differed for the three mouse strains: CF1, 0.24 +/- 0.08 mg; MTN, 0.92 +/- 0.22 mg; and MT1,2KO, 1.7 +/- 0.4 mg. Gene array results showed that nearly all arrayed genes were unaffected by cadmium. However, MT1 and MT2 had Cd+/Cd- expression ratios >1 in all four groups, while all ratios for MT3 were essentially 1, showing specificity. Both probes for MAPK 14 (p38 MAPK) had expression ratios >1, while no other MAPK responded to cadmium. Vacuolar proton pump ATPase and integrin alpha v (osteoclast genes), transferrin receptor, and src-like adaptor protein genes were stimulated by Cd; other src-related genes were unaffected. Genes for bone formation, stress response, growth factors, and signaling molecules showed little or no response to cadmium. Results support the hypothesis that Cd stimulates bone demineralization via a p38 MAPK pathway involving osteoclast activation.     

Cadmium in milk and mammary gland in rats and mice

The purpose of the present investigation was to study the uptake of cadmium in mammary tissue, effects on milk secretion and composition, and lactational transport of cadmium to the sucklings. Cadmium exposure during lactation resulted in retention of cadmium in the mammary tissue in mice and rats. The uptake of cadmium in the mammary tissue was rapid, as shown in lactating mice by whole-body autoradiography 4 h after an intravenous injection of a tracer dose of (109)CdCl(2). Retention of cadmium in kidneys of suckling pups was observed in the autoradiograms at 7 days after exposure of the dams. Lactating rats were intravenously infused with (109)CdCl(2) in 0.9% saline via osmotic minipumps from day 3 to day 16 after parturition. The cadmium dose given was 0, 8.8, 62 and 300 microg Cd/kg body wt. per day. Plasma and milk were collected at day 10 and 16 after parturition. Plasma cadmium levels in dams increased from day 10 to day 16. Cadmium levels were higher in milk than in plasma, with milk/plasma ratios varying from 2 to 6. Zinc levels in milk were positively correlated to cadmium levels in milk (r(2)=0.26; P=0. 03). In milk, (109)Cd was distributed in fat (46-52%), casein fraction (40-46%), and whey fraction (6-8%). There was a high correlation between cadmium concentrations in pups' kidney and cadmium concentrations in dam's milk (r(2)=0.98; P < 0.001). Of the cadmium dose given to the dams <0.05% was retained in the litters on day 16 of lactation. No effects were observed due to cadmium exposure on body weight in pups or dams. Cadmium treatment did not cause any effect on the lactose or protein concentration in milk, the concentrations of DNA, RNA or the ratio RNA/DNA in the mammary gland. Histological evaluation of mammary tissue did not reveal any abnormalities at any dose level. (109)Cd was bound to metallothionein in mammary tissue. The fraction of radiolabelled cadmium bound to metallothionein increased in a dose-dependent manner in both the liver (88-98%) and mammary tissue (57-80%). The present results indicate a low transfer of cadmium to the suckling pup, which might be due to binding of cadmium to metallothionein in the mammary tissue. However, during the susceptible developmental period even a low cadmium exposure may be of concern.     

Increased metallothionein in mouse liver, kidneys, and duodenum during lactation.

Lactation-induced increases in cadmium absorption and retention have been demonstrated in mid-lactating mice, but no systematic measurements of endogenous metal-binding protein concentrations during lactation have been reported. Using Cd/hemoglobin radioassay, this study detected significant increases in metallothionein (MT) concentrations in liver (4-fold), kidneys (2-fold), and duodenum (2-fold), but not jejunum, of mouse dams on days 13 and 20 of lactation. These increases occurred in the absence of cadmium exposure and were specific to the lactation period; dams 5 days after weaning showed MT levels that were similar to those of nonpregnant (NP) mice. Similarly, Northern blot analyses of livers from lactating mice demonstrated that MT mRNA concentrations in maternal liver during mid-lactation were 6-fold higher than those observed 5 days after pups were weaned. Gel filtration of final supernatants from the Cd/hemoglobin assay confirmed that the Cd-binding molecule induced during lactation was indeed metallothionein. In addition, chromatographic analyses of cytosols from tissues taken from dams administered small amounts of Cd (66 ng/mouse) showed that the trace amounts of Cd absorbed through the maternal gastrointestinal tract during mid-lactation were also bound to the MT. These results indicate MT induction in mouse dams occurs as a physiological consequence of lactation, requiring no external stimulus. This induced MT participates in binding low levels of dietary cadmium consumed by the dam. During lactation, elevated maternal MT may affect pathways for essential trace metals as well as sequester toxic metals harmful to the neonate. Multiparous humans may have increased risk of accumulating environmental Cd.     

Neonatal and Maternal Body Burdens of Hexachlorobenzene (HCB) in Mice: Gestational Exposure and Lactational Transfer

Neonatal and Maternal Body Burdens of Hexachlorobenzene (HCB)in Mice: Gestational Exposure and Lactational Transfer. COURTNEY,K. D., and ANDREWS, J. E. (1985). Fundam. Appl. Toxicol. 5,265–277. Hexachlorobenzene (HCB), a ubiquitous lipophilicpollutant, was readily transferred in the milk of lactatingdams to their suckling neonates. Pregnant CD-1 mice were treatedduring gestation, and the body burdens of HCB in the neonatesand the dams were determined during lactation. Also, neonatesfrom dams treated with HCB during gestation were cross-fosteredat birth to dams treated with corn oil during gestation. Thebody burdens of HCB were greater in the neonates exposed toHCB by lactational transfer than the neonates exposed only bygestational transfer. In many tissues, the concentration ofHCB in the pups from full litters was similar to that in pupsfrom litters reduced to two pups per litter. Lactational transferof HCB from the dams to the pups was a major route of excretionin that 95% of HCB was depleted during 20 days of lactation.HCB depletion was similar in dams with whole litters, and thosewith litters reduced to two pups.     

Pregnancy-induced mobilization of copper and zinc bound to renal metallothionein in cadmium-loaded rats

Our investigations undertook to examine whether copper (Cu) and zinc (Zn) bound to renal metallothionein (MT) along with cadmium (Cd) in Cd-loaded rats can be mobilized during pregnancy and lactation. Rats of the Wistar strain were injected with Cd. Synthesis of MT containing Cd, Cu and Zn was induced in the kidneys. Concentrations of Cd and essential elements in blood plasma and organs (liver and kidneys) were compared among non-pregnant and pregnant, and Cd-loaded and non-loaded rats at middle and late gestational days and after delivery. Cu bound to renal MT was decreased with gestational age, while Zn was slightly increased. The results indicate that Cu was mobilized and utilized even when the metal was bound to MT in the kidneys of dams. On the other hand, Zn bound to renal MT in Cd-loaded rats was retained and not mobilized during pregnancy. The elution profile of renal MT on an SW column was changed from a typical renal to a mixed profile of renal and hepatic MTs as a result of decreased Cu content in MT. Plasma essential elements changed similarly with gestational age in both concentrations and distributions in non-loaded and Cd-loaded rats. These results indicate that the 3 metals bound to renal MT are dealt with differently during gestation and lactation, and Cu but not Zn is transferred to the fetus, independently of the Cd status of the dam.     

Turnover of parenterally administered zinc and cadmium and the redistribution of metallothionein bound zinc in newborn rats

The whole body retention, tissue distribution and protein binding patterns of 65Zn were compared with 109Cd in newborn rats during postnatal development. One-day-old pups received a single injection of either 65Zn (2.5 microCi) or 109Cd (2.5 microCi plus 1 mg Cd/kg as CdCl2). During the 22 days of age, the whole body retention of 109Cd was higher than that for 65Zn. The biological half times were 466 and 46.3 days for 109Cd and 65Zn, respectively. There were marked differences in tissue deposition of these metals. Both liver and kidney accumulated more 109Cd than other tissues while the 65Zn showed a uniform distribution, with a gradual decrease in radioactivity with age. At the time of weaning, 109Cd had accumulated mainly in liver and kidney whereas, 65Zn was found predominantly in bone and skin. The specific binding of 109Cd to hepatic MT in newborn rats did not change with growth. Although a significant amount of 65Zn initially accumulated in the MT fractions in the liver, it was transferred gradually to high molecular weight protein fractions during development. The administration of these 2 metals had no effect on the body weight, liver weight and total hepatic zinc concentration. However, a significantly high content of MT and zinc in MT fractions was detected in the livers of Cd-treated rats at 22 days of age. The results show the transfer of the essential metal, zinc from hepatic MT to other proteins and the specific binding of cadmium, the non-essential metal to MT during postnatal development in rats.     

Gastrointestinal absorption of cadmium in mice during gestation and lactation: II. Continuous exposure studies

The effect on cadmium retention of continuous exposure to drinking water containing low levels of cadmium during pregnancy and lactation was studied in mice. Female mice were provided drinking water ad libitum containing ¹⁰⁹CdCl₂ (0.03 μCi ¹⁰⁹Cd/ml, 0.11 ppb total cadmium) throughout either gestation, lactation, or a combined period of pregnancy and lactation. Nonpregnant control mice were exposed to the same cadmium solution for similar time periods. Dams in all three experimental groups retained two to three times more cadmium (expressed as percentage of ingested dose) than did nonpregnant controls. The ¹⁰⁹Cd contents of liver, kidney, mammary tissue, and duodenum increased strikingly in all three groups. Increases in kidney and mammary tissue were particularly apparent during lactation, with increases of fivefold for kidney and at least ninefold for mammary tissue, compared to levels in nonpregnant controls. Increases in ¹⁰⁹Cd retention by the duodenum were fivefold during gestation and three- to fourfold during lactation. The kidneys of dams exposed during lactation retained 53% of the whole body ¹⁰⁹Cd, while kidneys of nonpregnant controls retained only 27%. Results indicate that pregnant and lactating mice absorb and subsequently retain substantially more cadmium from their diets than do nonpregnant mice.     

Prolactin lowering activity of the retinoid Ro 14-9706 affecting lactation and pup survival

The arotinoid Ro 14-9706, though devoid of any teratogenic potential, was found to reduce dose dependently the survival of pups when their mothers were treated with toxic doses during days 6 – 15 of gestation. The increased mortality was primarily seen during early lactation. When pups derived from treated mothers were nursed by control foster mothers unexposed to the drug, their survival was significantly improved indicating that the increased mortality was not solely due to fetal drug exposure. When pups derived from untreated mothers were fostered by dams that were exposed to the arotinoid during pregnancy, a significant pup mortality (p <0.01) was observed, suggesting that the nursing behaviour of lactating dams was seriously affected. This impairment could be linked to a prolactin-suppressive activity of the arotinoid during lactation which was also seen during pregnancy. Other pituitary hormones, however, were not affected by the compound. Although the drug induced pronounced structural alterations in mitochondria of adrenocortical cells, visualized by light microscopy as extended vacuolization in the zona fasciculata and reticularis, this pathological finding did not translate into functional impairment of steroidogenesis. Thus, the arotinoid Ro 14-9706 exhibits in rats a prolactin-suppressive activity which affects lactation and subsequently pup survival. This particular endocrinological interference is a new phenomenon and uncommon for retinoids. Received: 14 June 1993/Accepted: 11 January 1994     

Neonatal mortality from in utero exposure to perfluorooctanesulfonate (PFOS) in Sprague-Dawley rats: dose-response, and biochemical and pharamacokinetic parameters

Perfluorooctanesulfonate (PFOS) is a widely distributed, environmentally persistent acid found at low levels in human, wildlife, and environmental media samples. Neonatal mortality has been observed following PFOS exposure in a two-generation reproduction study in rats and after dosing pregnant rats and mice during gestation. Objectives of the current study were to better define the dose-response curve for neonatal mortality in rat pups born to PFOS-exposed dams and to investigate biochemical and pharmacokinetic parameters potentially related to the etiology of effects observed in neonatal rat pups. In the current study, additional doses of 0.8, 1.0, 1.2, and 2.0 mg/kg/day were included with original doses used in the two-generation study of 0.4 and 1.6 mg/kg/day in order to obtain data in the critical range of the dose-response curve. Biochemical parameters investigated in dams and litters included: (1) serum lipids, glucose, mevalonic acid, and thyroid hormones; (2) milk cholesterol; and (3) liver lipids. Pharmacokinetic parameters investigated included the interrelationship of administered oral dose of PFOS to maternal body burden of PFOS and the transfer of maternal body burden to the fetus in utero and pup during lactation, as these factors may affect neonatal toxicity. Dosing of dams occurred for 6 weeks prior to mating with untreated breeder males, through confirmed mating, gestation, and day four of lactation. Dose levels for the dose-response and etiological investigation were 0.0, 0.4, 0.8, 1.0, 1.2, 1.6, and 2.0 mg/kg/day PFOS. Statistically significant decreases in gestation length were observed in the 0.8 mg/kg and higher dose groups. Decreases in viability through lactation day 5 were observed in the 0.8 mg/kg and higher dose groups, becoming statistically significant in the 1.6 and 2.0 mg/kg dose groups. Reduced neonatal survival did not appear to be the result of reductions in lipids, glucose utilization, or thyroid hormones. The endpoints of gestation length and decreased viability were positively correlated, suggesting that late-stage fetal development may be affected in pups exposed to PFOS in utero and may contribute to the observed mortality. Benchmark dose (BMD) estimates for decreased gestation length, birth weight, pup weight on lactation day 5, pup weight gain through lactation day 5, and viability resulted in values ranging from 0.27 to 0.89mg/kg/day for the lower 95% confidence limit of the BMD5 (BMDL5). Results of analyses for PFOS in biological matrices indicate a linear proportionality of mean serum PFOS concentration to maternal administered dose prior to mating and through the first two trimesters of gestation. However, at 21 days of gestation, mean serum PFOS concentrations were notably reduced from values measured earlier in gestation. Urinary and fecal elimination was low as expected from prior observations in adult rats. Significant transfer of PFOS from dam to fetus in utero was confirmed, and results suggest that dam and corresponding fetal body burdens, as indicated by serum and liver PFOS levels, correlate with neonatal survival.     

Protective role of zinc against the neurotoxicity induced by exposure to cadmium during gestation and lactation periods on hippocampal volume of pups tested in early adulthood

The present study was conducted to assess the possible effect of cadmium (Cd) throughout gestation and lactation on the volume of the subregion of the hippocampus as well as the potential protective role of zinc (Zn) against Cd neurotoxicity. For this purpose, female rats received either tap water, Cd, Zn or Cd?+?Zn in their drinking water during gestation and lactation. At postnatal day 35 (PND35), the male pups were sacrificed, and their brains were taken for histologic, chemical, and biochemical analysis. Hippocampal volume was measured in histologic brain slices using Cavalieri’s principle. Zn depletion was observed in the brains of pups issued from mothers exposed to Cd. Biochemical analysis further revealed that Cd exposure significantly increases the superoxide dismutase (SOD) activity, as well as the metallothionein (MT) level. During histologic investigation, our results showed that gestational and lactational exposure to Cd significantly altered and decreased the volume of CA1, CA3 pyramidal cell layer and the dentate gyrus. However, there were no marked differences shown in CA2 subfield. Compared to Cd group, co-treatment with Cd and Zn provided correction of the changes induced by the Cd exposure. These results highlight the protective role of Zn against Cd-induced alteration in the hippocampus which is a crucial structure implicated in learning and memory processes.     

Behavioural effects of phenobarbitone II. Effects in nursing female mice

Phenobarbitone supplied at concentrations of 187.5 mg/l and 500 mg/l (50--60 mg/kg and 120--190 mg/kg daily) in the drinking fluid of mice throughout pregnancy and lactation did not affect gestation period, numbers born or resorbed or pup weights at birth and weaning. The higher dose caused significant retardation in eye-opening, development of forelimb extension, negative geotaxis, in appearance of washing and self-grooming behaviours and in cessation of suckling. The lower dose had no detectable effects. Ethological methods were used to examine spontaneous behaviour of nursing dams within home cages on days 1, 7, 14 and 21 postpartum. Treated dams showed differences in behaviour from controls, the most marked of which were increased levels of Maternal Behaviour and decreases in Non-Social Activity in the high-dose group at day 1, and continuance of pup Nursing in both treated groups and Nest Building in the high-dose group at day 21 when controls were ceasing these behaviours. At 21 days all treated dams also showed less Immobility than controls and high-dose dams a smaller amount of pup Social Investigation. Many behavioural differences at 21 days were related to developmental delay of treated pups. Overall, phenobarbitone did not impair maternal care.     

Computer Simulation of the Lactational Transfer of Tetrachloroethylene in Rats Using a Physiologically Based Model

A physiologically based pharmacokinetic (PBPK) model describing transfer of inhaled tetrachloroethylene (PCE) into breast milk with the consequent exposure of the nursing pups was developed and validated in lactating rats. The PBPK model was based on partition coefficients (PCs), which were determined by the equilibration of tissues with known concentrations of PCE vapor. The blood/air and milk/blood PCs for the dam were 33.5 and 12.0, the liver/blood and fat/blood PCs for the dam were 1.9 and 42.4, and the slowly perfused tissues/blood and rapidly perfused tissues/blood PCs for the dam were 0.9 and 1.7, respectively. The blood/air PC for the pup was 24.3, and the solid tissues/blood PC for the pup was 4.5. Metabolic constants were determined by a gas uptake method. The pseudo-maximal velocity of PCE metabolism in the dam was low, 0.03 mg/kg/hr, and the apparent Michaelis-Menten constant was 0.3 mg/liter. The lactating Sprague-Dawley females were exposed to PCE vapor for 2 hr with concentrations ranging from 20 to 1000 ppm PCE. The dams were returned to the pups after exposure and the pups received PCE via breast milk while they nursed. PCE concentrations in the air, blood, milk, and tissues were determined by gas chromatography and compared to PBPK model predictions. Nursing resulted in a peak pup blood PCE concentration of 0.9 mg/liter and a peak pup solid tissue PCE concentration of 8 mg/kg. The PBPK lactation model adequately described the distribution of inhaled PCE in maternal blood and milk, as well as in the nursed pup′s gastrointestinal tract, blood, and solid tissues. It is concluded that the predictions based on PBPK modeling for disposition of PCE in lactating rats and their pups were in good agreement with the experimental data.     

Transplacental and Lactational Exposure to Mercury in Hamster Pups after Maternal Administration of Methyl Mercury in Late Gestation

Abstract: Pregnant Syrian golden hamsters were given a single oral dose of ²⁰³Hg-labelled methyl mercury (MeHg), 1.6 μmol/kg body weight, on day 12 of gestation. The uptake, retention and tissue distribution of ²⁰³Hg in the dams and pups was studied by γ-counting during the following three weeks. The average transplacental transfer of ²⁰³Hg was 1.1% of the administered dose per pup, corresponding to 11% of the administered dose to a whole litter. This was considerably more than in our previous studies when the dams were treated on gestational day 2 (1.3%) or 9 (4.6%). The amount of ²⁰³Hg transferred to each pup in utero was independent of the litter size. The average additional transfer of ²⁰³Hg to a litter via milk was 1.7% of the administered dose. In the pups, the content of ²⁰³Hg in the liver and brain decreased, while the content in the kidneys and pelt increased during the second and third week. The highest amount of ²⁰³Hg was generally found in the pelt, which indicated that unweaned hamster pups primarily excrete MeHg by binding to hair. The chemical form of mercury in the liver and kidneys of the pups was determined by ion-exchange separation of inorganic Hg and MeHg followed by γ-counting. The amount of inorganic Hg in the liver of the pups remained constant throughout the experiment, while it increased in the kidneys after one week due to the demethylation of MeHg. The inorganic Hg in the liver of newborn pups was probably due to maternal demethylation of MeHg and transplacental transfer of inorganic Hg. This hypothesis was supported by the observed transplacental transfer of inorganic Hg in pregnant dams administered ²⁰³HgCl₂ on day 12 of gestation.     

Effects of daily interrupted nursing on body weight and survival of Fischer 344 rat pups

The effects on Fischer 344 rat pup body weight and survival were investigated when the dam was separated from her litter and deprived of food and water, approximately 7 h per day, from day 5 to day 21 of the postnatal period. This separation procedure simulates an inhalation reproduction study in which only the dams are exposed to the test material during lactation. The dams and pups were observed daily for clinical signs, and the number of pups in each litter and their body weights were determined on days 4, 9, 14 and 21 after parturition. When pups and dams were so separated, there was no effect on pup survival, but the pup body weights were moderately lower for this group by the end of the postnatal observation period.     

The influence of maternal cadmium exposure or fetal cadmium injection on hepatic metallothionein concentrations in the fetal rat

The ability of Cd to induce the synthesis of fetal hepatic metallothionein (MT) was investigated in rat fetuses exposed to Cd throughout gestation via the mother's drinking water or injected directly with Cd through the uterine wall on Day 18 of gestation. On Day 21 all dams were killed and fetal and maternal tissues were removed. Tissue MT, Zn, Cu, and Cd concentrations were measured. Fetal hepatic Cd concentration was increased only at the high maternal Cd exposure, whereas Zn concentration was significantly reduced by Cd exposure. Both fetal liver and kidney MT were reduced following maternal Cd exposure. Unlike maternal hepatic MT, fetal hepatic MT was not increased after maternal Cd exposure nor did the direct injection of Cd into the 18 days of gestation fetus induce fetal MT synthesis. These data suggest that fetal rat liver is incapable of synthesizing MT in response to Cd, possibly because Cd is not transported to the site of MT synthesis in the fetal system. Furthermore, neither the route of exposure, the duration of prenatal Cd exposure, nor the stage of gestation appear to account for the differences observed between fetal and adult hepatic MT induction by Cd.     

Copper-carbohydrate interaction in maternal, fetal and neonate rat

The present study was undertaken to determine whether the same type of interaction between dietary fructose and copper that affects young growing male rats also affects the fetus and the neonate. Female rats were fed copper-deficient (0.6 μg Cu/g) or adequate (6.0 μg/g) diets containing 62% carbohydrate as fructose or starch either for 8 weeks prior to conception, and during mating, gestation and lactation, or just during gestation. Fetuses were killed at days 14, 18 or 21 of gestation and newborn pups were killed at days 0, 10, or 21 postpartum. Regardless of the duration of dietary copper deprivation, feeding the fructose diet deficient in copper during pregnancy resulted in either fetal resorption or mortality of all newborn pups during the first few hours postpartum. In contrast, copper-deficient rats fed the starch-containing diets delivered live pups. However, 40% of their pups died during the first 2 days postpartum and occurred only when dams had been fed the deficient diet for 12–13 weeks. When fed the deficient diet for a total of 3 weeks only, during pregnancy, all copper-deficient rats fed starch delivered live pups and no mortality occurred during the lactation period. Feeding the copper-adequate fructose diet during lactation resulted in a lower hepatic copper concentration of suckling pups compared with starch feeding. Female pups had higher levels of copper and iron than male pups. The data show that fetal resorption and mortality of the neonate pup was dependent on the type of dietary carbohydrate fed to copper-deficient animals during pregnancy.     

Lack of differential sensitivity to cholinesterase inhibition in fetuses and neonates compared to dams treated perinatally with chlorpyrifos.

Pregnant Sprague-Dawley rats were exposed to chlorpyrifos (CPF; O,O-diethyl-O-[3,5,6-trichloro-2-pyridinyl] phosphorothioate) by gavage (in corn oil) from gestation day (GD) 6 to postnatal day (PND) 10. Dosages to the dams were 0 (control), 0.3 (low), 1.0 (middle) or 5.0 mg/kg/day (high). On GD 20 (4 h post gavage), the blood CPF concentration in fetuses was about one half the level found in their dams (high-dose fetuses 46 ng/g; high-dose dams 109 ng/g). CPF-oxon was detected only once; high-dose fetuses had a blood level of about 1 ng/g. Although no blood CPF could be detected (limit of quantitation 0.7 ng/g) in dams given 0.3 mg/kg/ day, these dams had significant inhibition of plasma and red blood cell (RBC) ChE. In contrast, fetuses of dams given 1 mg/kg/day had a blood CPF level of about 1.1 ng/g, but had no inhibition of ChE of any tissue. Thus, based on blood CPF levels, fetuses had less cholinesterase (ChE) inhibition than dams. Inhibition of ChE occurred at all dosage levels in dams, but only at the high-dose level in pups. At the high dosage, ChE inhibition was greater in dams than in pups, and the relative degree of inhibition was RBC approximately plasma > or = heart > brain (least inhibited). Milk CPF concentrations were up to 200 times those in blood, and pup exposure via milk from dams given 5 mg/kg/day was estimated to be 0.12 mg/kg/day. Therefore, the dosage to nursing pups was much reduced compared to the dams exposure. In spite of exposure via milk, the ChE levels of all tissues of high-dosage pups rapidly returned to near control levels by PND 5.     

Refining the effects observed in a developmental neurobehavioral study of ammonium perchlorate administered orally in drinking water to rats. I. Thyroid and reproductive effects

A recent study further investigated the potential effects of maternal thyroid function and morphology on fetal development upon maternal exposure to ammonium perchlorate during gestation and lactation. Female Sprague-Dawley rats (25/group) were given continual access to 0 (carrier), 0.01, 0.1, 1.0, and 30.0 mg/kg-day perchlorate in drinking water beginning 2 weeks prior to cohabitation through lactation day 10. Maternal, fetal, and pup serum thyroid hormone (thyroid-stimulating hormone [TSH], triiodo thyronine [T(3)], thyroxine [T(4)]) levels and thyroid histopathology were evaluated on gestation day 21, and lactation days 5, 10, and 22. No effects of exposure were observed on cesarean-sectioning, litter parameters, or fetal alterations. Reproductive parameters, including gestation length, number of implants, litter size, pup viability, and lactation indices, were comparable among all groups. Thyroid weights of dams sacrificed on gestation day 21, and lactation days 10 and 22 were significantly increased at 30.0 mg/kg-day. Increased thyroid weights were observed in male and female pups as early as postpartum days 5 and 10, respectively. Changes in maternal and neonatal thyroid histopathology were detectable at 1.0 mg/kg-day exposure. The maternal no-observable-effect level (NOEL) was 0.1 mg/kg-day (follicular cell hyperplasia was present at 1.0 and 30.0 mg/kg-day). The developmental NOEL was less than 0.01 mg/kg-day; thyroid weights of postpartum day 10 pups were increased at all exposures. Colloid depletion at 1.0 and 30.0 mg/kg-day exposures and changes of hormone levels at all exposures were considered an adaptive effect and appeared reversible in the rodent.     

Developmental toxicity of perfluorooctane sulfonate (PFOS) is not dependent on expression of peroxisome proliferator activated receptor-alpha (PPAR alpha) in the mouse

Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) are members of a family of perfluorinated compounds. Both are environmentally persistent and found in the serum of wildlife and humans. PFOS and PFOA are developmentally toxic in laboratory rodents. Exposure to these chemicals in utero delays development and reduces postnatal survival and growth. Exposure to PFOS on the last 4 days of gestation in the rat is sufficient to reduce neonatal survival. PFOS and PFOA are weak agonists of peroxisome proliferator activated receptor-alpha (PPAR alpha). The reduced postnatal survival of neonatal mice exposed to PFOA was recently shown to depend on expression of PPAR alpha. This study used PPAR alpha knockout (KO) and 129S1/SvlmJ wild type (WT) mice to determine if PPAR alpha expression is required for the developmental toxicity of PFOS. After mating overnight, the next day was designated gestation day (GD) 0. WT females were weighed and dosed orally from GD15 to 18 with 0.5% Tween-20, 4.5, 6.5, 8.5, or 10.5mg PFOS/kg/day. KO females were dosed with 0.5% Tween-20, 8.5 or 10.5mg PFOS/kg/day. Dams and pups were observed daily and pups were weighed on postnatal day (PND) 1 and PND15. Eye opening was recorded from PND12 to 15. Dams and pups were killed on PND15, body and liver weights recorded, and serum collected. PFOS did not affect maternal weight gain or body or liver weights of the dams on PND15. Neonatal survival (PND1-15) was significantly reduced by PFOS in both WT and KO litters at all doses. WT and KO pup birth weight and weight gain from PND1 to 15 were not significantly affected by PFOS exposure. Relative liver weight of WT and KO pups was significantly increased by the 10.5mg/kg dose. Eye opening of PFOS-exposed pups was slightly delayed in WT and KO on PND13 or 14, respectively. Because results in WT and KO were comparable, it is concluded that PFOS-induced neonatal lethality and delayed eye opening are not dependent on activation of PPAR alpha.     

Fatty acid alterations in liver and milk of cadmium exposed rats and in brain of their suckling offspring

Fatty acid composition was studied in milk at day 14 and in liver at day 24 after parturition of lactating rats exposed to 0 ppm, 5 ppm or 25 ppm cadmium (Cd) via drinking water for 17 days during lactation, and in the brain of their offspring at day 19 after birth. In the liver phospholipid fraction, 22:5(n-3) was significantly higher, while in the triacylglycerol fraction 22:6(n-3)/20:5(n-3) ratio was significantly lower in the 25 ppm group compared to the controls. Significantly higher proportions of 16:0 and lower proportions of medium-chain fatty acids, 8:0-14:0, were observed in milk of dams in the 25 ppm group, indicating decreased enzymatic activity of thiotransferase II in the mammary gland. Slightly increased levels of 20:3(n-6) were observed in brains of pups in the 25 ppm group compared to control. The results indicate that Cd exposure influences fatty acid metabolism in lactating rats.