Follicular Cell Height Research Articles

Reproductive and Developmental Toxicity of a Pentabrominated Diphenyl Ether Mixture, DE-71, to Ranch Mink (Mustela vison) and Hazard Assessment for Wild Mink in the Great Lakes Region

Polybrominated diphenyl ethers (PBDEs) are widespread persistent and bioaccumulative environmental contaminants. Recent scientific attention has focused on the developmental toxicity of PBDE commercial mixtures following perinatal exposure of rodents; however, these studies do not necessarily predict toxicity to highly exposed top predators, such as mink (Mustela vison). Here we assessed the effects of environmentally relevant doses (0, 0.1, 0.5, and 2.5 ppm [wt/wt] in feed) of a technical pentabrominated diphenyl ether mixture, DE-71, on reproductive performance of mink and on development of offspring exposed perinatally and post-weaning until 33 weeks. A dietary concentration that causes no effects on reproduction in rodents, 2.5-ppm DE-71, resulted in complete reproductive failure in these mink, while whelping rates were not affected at all lower does. Developmental effects in offspring were evident in 33-week-old juveniles, which were more sensitive to effects than their respective dams. Juvenile thyroid hormone homeostasis was also much more sensitive compared to rodents, and at 0.5-ppm DE-71, total triiodothyronine (T3) was significantly decreased in all males and females, even despite a compensatory increase of total thyroxine (T4) in females. T4-outer-ring deiodinase activity, mainly contributed by type II deiodinase, was not affected at any dose for any life stage, but thyroid follicular epithelium cell height was elevated in the 0.5-ppm-treated juveniles (p = 0.057). Ethoxyresorufin O-deethylase activity was significantly induced in all offspring at 33 weeks, most likely as a consequence of polybrominated dioxin, furan, or biphenyl impurities in DE-71. Biomonitoring of wild mink in the Great Lakes region indicated that most populations had lower concentrations than what are expected to affect thyroid hormone homeostasis, but margins of safety are small and mink around Hamilton Harbour exceeded the no observed adverse effect level for T3 disruption.

The impact of the goitrogen 6-propylthiouracil (PTU) on West-African clawed frog ( Xenopus tropicalis) exposed during metamorphosis

This study investigated the suitability of using tadpoles of the West-African clawed frog ( Xenopus tropicalis) for studying adverse effects on the thyroid hormone system after chemical exposure. Tadpoles were exposed to the thyroxine synthesis inhibitor 6-propylthiouracil (PTU) at concentrations between 2–75 mg/L during 14 days. After 5 and 14 days of exposure the developmental stage, hind limb length, body length and weight were measured. Moreover, histological measurements of the thyroid glands were performed after 14 days of exposure. These measurements included maximum thyroid cross-section area, follicular area and epithelial cell height. Tadpoles in the 75 mg/L treatment were less developed and had shorter hind limb length than the control group after 14 days of exposure. No effects were recorded on these parameters at lower PTU concentrations. The histological measurements revealed clear dose–response relationships in both follicular cross-section area and epithelial cell height, with lowest observed effect concentrations (LOECs) recorded at 2 and 5 mg/L, respectively. This study shows that X. tropicalis is a suitable species for detection of thyroid disrupting chemicals. Further, histopathological measurements of thyroid glands are more sensitive parameters compared with apical endpoints when studying adverse effects on thyroid hormone system caused by PTU exposure in X. tropicalis.

Evaluation of ammonium perchlorate in the endocrine disruptor screening and testing program's male pubertal protocol: Ability to detect effects on thyroid endpoints

The U.S. EPA Endocrine Disruptor Screening Program (EDSP) Tier 1 male pubertal protocol was designed as a screen to detect endocrine-disrupting chemicals which may alter reproductive development or thyroid function. One purpose of this in vivo screening protocol is to detect thyrotoxicants via a number of different mechanisms of action, such as thyroid hormone synthesis or clearance. Here we evaluate the ability of this EDSP male pubertal protocol to detect the known thyrotoxicant ammonium perchlorate as an endocrine disruptor. Ammonium perchlorate is a primary ingredient in rocket fuel, fertilizers, paints, and lubricants. Over the past 50 years, potassium perchlorate has been used to treat hyperthyroidism in humans. Perchlorate alters thyroid hormone secretion by competitively inhibiting iodide uptake by the thyroid gland. In this study, ammonium perchlorate was administered at 62.5, 125, 250, and 500 mg/kg to male Wistar rats based on a pilot study of oral dosing. Doses of 125–500 mg/kg perchlorate decreased T4 in a dose-dependent manner. TSH was significantly increased in a dose-responsive manner at the same doses, while T3 was unchanged at any dose. Thyroid histology was significantly altered at all doses, even at the 62.5 mg/kg, with a clear dose-dependent decrease in colloid area and increase in follicular cell height. No effects on preputial separation, a marker of pubertal progression, or reproductive tract development were observed at any dose. These results demonstrate that the male pubertal protocol is useful for detecting thyrotoxicants which target the thyroid axis by this mechanism (altered uptake of iodide). This study also found that perchlorate exposure during this period did not alter any of the reproductive developmental endpoints.

Thyrotoxicity of Sodium Arsenate, Sodium Perchlorate, and Their Mixture in Zebrafish Danio rerio

Both perchlorate and arsenate are environmental contaminants. Perchlorate is a definitive thyroid disruptor, and arsenic may disrupt thyroid homeostasis via multiple pathways. To evaluate the effects of sodium perchlorate and sodium arsenate on thyroid function and possible interactions between them, zebrafish (Danio rerio) were exposed to sodium perchlorate (10 and 100 mg/L), sodium arsenate (1 and 10 mg/L), and the mixture sodium perchlorate + sodium arsenate (10 + 1 and 100 + 10 mg/ L) for up to 90 days. At day 10, 30, 60, and 90, fish were sampled and analyzed forthyroid histopathological end points including follicular cell height, follicle size, colloid size, colloid depletion, hyperplasia, and angiogenesis. Effects on epithelial cell height (hypertrophy) were seen as early as 10 days after exposure. Perchlorate induced changes in all parameters staring at 30 days of exposure. Prolonged perchlorate exposure induced angiogenesis, a relatively new marker of thyroid disruption. Sodium arsenate was less effective than sodium perchlorate in causing thyroid histopathologies, but transient responses were seen for hypertrophy, hyperplasia, and colloid depletion (% colloid). This is the first report of arsenate-induced effects on thyroid histopathology. However, because statistically significant effects were not consistently seen in all end points, evidence for arsenate as a thyroid disruptor remains equivocal. In general, the sensitivity of the following histopathological indicators for indicating thyroid perturbations is, in descending order: follicular cell height > percent of colloid area/follicle area > colloid area/follicular cell height > hyperplasia > angiogenesis > colloid area >follicle area = fish growth.

EFFECT OF CONJUGATED EQUINE ESTROGENS AND TAMOXIFEN ADMINISTRATION ON THYROID GLAND HISTOMORPHOLOGY OF THE RAT

The aim of this study was to evaluate the action of the conjugated equine estrogens and tamoxifen on the morphology of thyroid gland in ovariectomized (OVx) rats. Conjugated equine estrogens (CEE), clinically used as estrogen therapy, is a complex formulation containing multiple estrogens that decrease menopausal symptoms. Thirty ovariectomized rats were randomly divided into 3 treatment groups: GI, vehicle (propylene glycol); GII, CEE 200 microg/kg per day; and GIII, tamoxifen 1 mg/kg per day. Another group of 10 rats with intact ovaries (GIV) was included, treated with the vehicle, and sacrificed during estrous. All animals were treated by gavage for 50 days, after which they were sacrificed. Blood samples were collected, and the thyroid was removed for morphological analysis and PCNA evaluation through immunohistochemical study. The thyroid follicular cell height was increased in animals treated with CEE (14.90 +/- 0.20 microm), with TAM (14.90 +/- 0.10 microm), and in rats with intact ovaries (15.10 +/- 0.50 microm) in comparison to that of the vehicle group (9.90 +/- 0.20 microm) (P < 0.001). The follicular area was larger in the CEE (2,225 +/- 51 microm2) and TAM (2,127 +/- 67 microm2) groups compared to that of the vehicle group (5,016 +/- 53 microm2). The levels of T4 and T3 in rats treated with CEE, with Tamoxifen and in rats with intact ovaries, were higher than those those in the vehicle group (P < 0.001). The PCNA index in the vehicle group was lower than in other groups. Our data suggest that estrogen and tamoxifen administration has a proliferative effect on the thyroid.

Perchlorate Affects Thyroid Function in Eastern Mosquitofish (Gambusia holbrooki) at Environmentally Relevant Concentrations

The purpose of this study was to determine the effects of perchlorate on thyroid function in mosquitofish. Adult mosquitofish were exposed to 0, 0.1, 1, 10, 100, and 1000 mg/L sodium perchlorate for 2, 10, and 30 d. Whole body thyroxin (T4) content and histological assessment of thyroid follicles (e.g., follicular epithelial height, hyperplasia, hypertrophy, and colloid depletion) were used to gauge alterations in thyroid function. Follicular epithelial cell height, hyperplasia, and hypertrophy increased with increasing perchlorate concentration, especially in fish exposed for 30 d, and these effects were statistically significantly different from control at concentrations as low as 0.1 mg/L (nominal concentration). The percent occurrence of follicles with depleted colloid decreased with increasing perchlorate concentration, which is contrary to what is expected with thyroid inhibition. There also was a decrease in whole body T4 concentration in fish exposed to perchlorate for 30 d, but clear dose-response relationships were less evident for whole body T4 than for histopathological endpoints. In conclusion, thyroid histopathology provides a sensitive biomarker for thyroid endocrine disruption at environmentally relevant concentrations of sodium perchlorate, and whole body T4 is a less sensitive indicator of perchlorate exposure than is histopathology.

The effect of cervical X-irradiation on activity index of thyrocytes and plasma TSH: A pre-clinical model for radiation-induced thyroid damage

Because radiotherapy in the head and neck region is necessary in the treatment of childhood cancer, possibilities to prevent damage to the thyroid gland must be explored. We developed a model in which radiation-induced effects can be investigated in a way that these effects can be quantified, using thyroid dysmorphology and plasma TSH. Thirty-five Wistar rats, 5 weeks old, were X-irradiated on the cervical region, with a single dose varying from 0 to 20 Gy. After 6 weeks, TSH, T4 and T3 were determined, and thyroid glands were processed for histological examination by two independent pathologists. A histological classification scale was developed, using follicular size, colloid density and cell height of thyrocytes to measure hyperplasia and hypertrophy. By the sum of these scores, a cell-activity index was calculated, which was related to plasma TSH concentration. Numbers of PAS-positive droplets and epithelial desquamation were also counted. Inter-observer reliability was assessed. Good to very good reliability was found for scores of follicular size, colloid density and cell height. Significant increase of cell-activity index was found after 10, 15 and 20 Gy. The plasma TSH concentration was positively correlated to the cell-activity index, increasing with radiation-doses up to 15 Gy. The number of desquamated cells was significantly increased after radiation doses >10 Gy, with moderate reliability. In conclusion, this model using cell-activity index of thyrocytes together with plasma thyrotropin concentrations and desquamation of cells can be used for interpretation and future (pre-clinical) studies of prevention of radiation-induced thyroid damage.

Thyroid Function in Nestling Tree Swallows and Eastern Bluebirds Exposed to Non-Persistent Pesticides and p, p?-DDE in Apple Orchards of Southern Ontario, Canada

This study examined the associated effects of pesticides and persistent residues of p,p'-DDE on thyroid function in 16-day-old tree swallows (Tachycineta bicolor) and 12-day-old eastern bluebirds (Sialia sialis). Apple orchards sprayed with pesticides in current use and reference sites were chosen for study in southern Ontario, Canada, during 2000-2001. We assessed thyroid hormone concentrations (plasma and muscle thyroxine [T4], and triiodothyronine [T3]) and the response to a challenge with thyroid stimulating hormone (TSH) as well as parameters of thyroid histology. Individual nests were exposed to as many as seven individual pesticide applications and up to five pesticide mixtures. Concentrations of p,p'-DDE in eggs from both species were highest in orchard sites, and ranged from 0.05 to 5.44 mg/kg wet weight for tree swallows, and from 0.17 to 95.4 mg/kg for eastern bluebirds. In 2000, tree swallows from sprayed orchards had elevated plasma T4 concentrations, and thyroid glands with collapsed follicles, hypertrophic epithelia, and higher follicular epithelial cells relative to chicks from reference sites. Plasma T4 concentration was positively correlated with the total number of pesticide mixtures applied during egg incubation through chick rearing. In 2001, basal plasma T3 concentration in tree swallow chicks did not differ between orchard and reference sites, though there were differences among orchards; T3 was positively correlated with total number of individual sprays applied during egg incubation through chick rearing. Tree swallows challenged with TSH showed no significant difference in thyroid hormone concentration or follicular epithelial cell height between exposure groups; however, a significantly greater proportion of chicks from reference sites were found to have thyroids with focal or diffuse hypertrophic epithelia relative to orchard chicks. In 2001, bluebirds from sprayed orchards had significantly higher T3 concentrations in response to TSH challenge; this response was positively correlated with the total number of pesticides applied during egg incubation through chick rearing.

Effects of ammonium perchlorate on thyroid function in developing fathead minnows, Pimephales promelas.

Perchlorate is a known environmental contaminant, largely due to widespread military use as a propellant. Perchlorate acts pharmacologically as a competitive inhibitor of thyroidal iodide uptake in mammals, but the impacts of perchlorate contamination in aquatic ecosystems and, in particular, the effects on fish are unclear. Our studies aimed to investigate the effects of concentrations of ammonium perchlorate that can occur in the environment (1, 10, and 100 mg/L) on the development of fathead minnows, Pimephales promelas. For these studies, exposures started with embryos of < 24-hr postfertilization and were terminated after 28 days. Serial sectioning of thyroid follicles showed thyroid hyperplasia with increased follicular epithelial cell height and reduced colloid in all groups of fish that had been exposed to perchlorate for 28 days, compared with control fish. Whole-body thyroxine (T4) content (a measure of total circulating T4) in fish exposed to 100 mg/L perchlorate was elevated compared with the T4 content of control fish, but 3,5,3′-triiodothyronine (T3) content was not significantly affected in any exposure group. Despite the apparent regulation of T3, after 28 days of exposure to ammonium perchlorate, fish exposed to the two higher levels (10 and 100 mg/L) were developmentally retarded, with a lack of scales and poor pigmentation, and significantly lower wet weight and standard length than were control fish. Our study indicates that environmental levels of ammonium perchlorate affect thyroid function in fish and that in the early life stages these effects may be associated with developmental retardation.

Effects of butylated hydroxyanisole on the development and functions of reproductive system in rats

Butylated hydroxyanisol (BHA) is a widely used antioxidant for long preservation of food products, cosmetics and pharmaceuticals. Although BHA is generally recognized as safe, it is classified as a suspected endocrine-disrupting compound. We investigated the effects of BHA on reproductive function and development by the treatment of mature male and female SD rats (F0) through pre-gestation, gestation and lactation period and of their offspring (F1) until 13 weeks old via gavage with BHA 0 (corn oil, vehicle control), 10, 100 and 500 mg/kg bw/day. Organ weights of liver, adrenal gland and thyroid gland of F0 rats were increased by BHA 500 mg/kg but those of spleen and ventral prostate were decreased without significant difference in terminal body weight. Reduced serum testosterone and thyroxine (T4) were observed with dose-dependent manner in F0 male rats. Mating rate was decreased and cohabitation duration for conception was longer without differences in the number, motility and morphology of sperm by BHA 500 mg/kg. Body weight of F1 offspring was significantly decreased with change of relative weight of liver and brain by BHA 500 mg/kg at PND21. Sexual maturation indicated by vaginal opening and preputial separation was delayed by BHA 500 mg/kg. The weights of liver and adrenal gland were increased while those of spleen, vagina, testes and ventral prostate were decreased in F1 rats exposed to BHA 100 or 500 mg/kg for 13 weeks. Also, BHA 500 mg/kg reduced the velocity of sperm motion and number with smaller-sized sperm head in F1 male rats and slightly shortened estrous cycle length with higher frequency of estrus and lower frequency of diestrus stages in F1 female rats. Lower serum T4 and testosterone contents with higher serum cholesterol levels were also observed by BHA 500 mg/kg. Increased follicular cell height, and exfoliated and vacuolated follicular epithelial cells were observed in thyroids of F1 female and males rats exposed to BHA 500 mg/kg. This study elucidates that high dose of BHA induce weak dysfunction and underdevelopment of reproductive system of male and female rats with the change of T4 and testosterone levels, sex organ weights and sexual maturation and histological lesions of thyroid gland.

Histological and histomorphometric alterations in thyroid and adrenals of CD rat pups exposed in utero to methyl thiophanate

Pregnant CD rats were treated with an initial dose of 0, 310 or 560 mg/kg bw per day of the fungicide methyl thiophanate (MT) on gestational days 10–14, corresponding to formation of thyroid and adrenal primordia; newborns were sacrificed on postnatal days (PNDs) 10 and 23. No apparent maternal toxicity and no effects on litter size, viability or weight gain were present. Delayed ear pinna detachment and eye opening were present at top dose level. Thyroid histology showed increased irregular nuclei and/or mitoses (PND 10—both doses), cells with necrotic or hydropic changes (PND 23—top dose). The adrenal cortex showed increased karyomegaly and hydropic degeneration (PND 23—both doses). Thyroid histomorphometry showed reduced follicular density, moderately increased follicular cell height and number of nuclei/follicle (PND 10—top dose and PND 23—both doses), suggesting retarded follicular maturation. The adrenal cortex relative area was slightly decreased (PND 10—top dose and PND 23—both doses). MT may act as weak endocrine disrupter, suggesting that attention should be paid to delayed endocrine alterations elicited by agrochemicals.

The herbicide metolachlor induces liver cytochrome P450s 2B1/2 and 3A1/2, but not thyroxine-uridine dinucleotide phosphate glucuronosyltransferase and associated thyroid gland activity.

Metolachlor (2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl) acetamide) is widely used internationally as a corn and cotton herbicide. The metolachlor effects noted in rats during testing for U.S. pesticide registration include increased liver weight and hepatocarcinogenicity associated with eosinophilic foci. These properties, plus nongenotoxicity, are also characteristic of the prototypical rat liver tumor promoter, phenobarbital. Phenobarbital induces hepatic cytochrome P450s CYP2B1/2 and CYP3A1/2 and thyroxine (T(4))-UDP-glucuronosyltransferase (T(4)-UGT), which enhances thyroxine clearance and thus indirectly increases thyroid gland activity. Because other chloroacetanilide herbicides are known to similarly affect rat thyroid gland, this study tested the hypothesis that metolachlor would have these additional phenobarbital-like effects on liver, especially that of T(4)-UGT induction with consequential stimulation of thyroid gland. Effects of metolachlor, fed to male Sprague-Dawley rats for 14 days at the carcinogenic dose of 3000 ppm, were compared to those of equimolar phenobarbital. Liver microsomal CYP2B1/2 and CYP3A1/2 were probed by immunoblotting and T(4)-UGT was measured enzymatically. Serum T(4), triiodothyronine (T(3)), and thyroid-stimulating hormone (TSH) and thyroid follicular epithelial cell morphology and proliferation were used to assess thyroid gland activity. Metolachlor induced CYP2B1/2 and CYP3A1/2 proteins, but unlike phenobarbital, did not affect T(4)-UGT activity. In agreement, serum T(4), T(3), or TSH were unaffected by metolachlor. Also, no significant effects of metolachlor on thyroid gland morphology or follicular epithelial cell height or proliferation were observed. These data demonstrate that metolachlor is an inducer of hepatic CYP2B1/2 activity. But unlike the prototypical CYP2B1/2 inducer phenobarbital, metolachlor does not cause an increase in T(4)-glucuronidation and thyroid gland activation.

Effects of maternal exposure to diethylstilbestrol on the development of the reproductive system and thyroid function in male and female rat offspring.

Diethylstilbestrol (DES) was administered subcutaneously at 1.5 or 15 microg/kg/day (DES 1.5 group, DES 15 group) to pregnant SD rats daily on days 7-21 of gestation to investigate its effects on the development and functions of the reproductive system and thyroid gland in their offspring. Of the 11 pregnant rats in the DES 15 group, only one delivered a live pup. Rat pups in the DES 1.5 group were autopsied at 1, 3, or 6 weeks after birth. In the DES 1.5 group, the plasma T4 concentrations at all weeks of age at autopsy were significantly increased, the TSH concentration at 6 weeks of age was also significantly increased, and the height of thyroid follicular epithelial cells was increased at 3 weeks. The testosterone concentration in the DES 1.5 group at 6 weeks was significantly decreased and the plasma LH concentration was increased. The DES treatment increased the plasma FSH concentration in female pups at 3 weeks, increased the percentages of primary and secondary ovarian follicles, and decreased the percentage of primordial follicles, but did not influence the timing of the vaginal opening or the onset of the estrous cycle. These observations indicate that prenatally administered DES increases thyroid function, and has an inhibitory effect on testicular function and a promoting effect on female reproductive function.

Morphological and functional differentiation of human thyroid cells in collagen gel culture

Objective: cultured human thyroid cells in collagen gel culture were examined on cell morphology and the production of thyroglobulin (Tg), triiodothyronine (T3) and thyroxine (T4) which are components of functional differentiation. Methods: thyroid cells obtained from normal human thyroid tissues (four cases), follicular adenoma tissues (three cases), papillary carcinoma tissues (three cases), and follicular carcinoma tissue (one case), were cultured in collagen gel. Then these cultured cells were observed on cellular morphology and production of Tg, T3 and T4. Moreover, changes in morphological characteristics and production of Tg, T3 and T4 induced by addition of thyroid stimulating hormone (TSH) and epidermal growth factor (EGF) to medium in collagen gel culture were determined. Results: normal and tumor cells in collagen gel culture formed colonies and follicles with Tg production, similar to in vivo-like three-dimensional cellular structures and functions. Normal thyroid cells stimulated TSH induced more Tg and produced morphological changes, i.e. enlarged follicular lumens and increased the height of follicular cells, but did not promote cell proliferation. Reversely, normal thyroid cells stimulated with EGF promoted cell proliferation, but did not change morphological findings and did not increase production of Tg, T3 and T4. Conclusion: these findings suggest that collagen gel culture is useful for observing the effects of stimulation by cell growth factor on the morphological and functional differentiation of human thyroid cells.

Cold-induced changes in thyroid function in a poikilothermic mammal, the naked mole-rat.

Cold acclimation induces very divergent responses in thyroid function in reptiles and mammals reflective of their different thermoregulatory modes. Naked mole-rats, unlike other small mammals, are unable to effectively employ endothermy and are operatively poikilotherms. We therefore investigated changes in their thyroid status with chronic cold exposure. Under simulated burrow conditions, free thyroxine (T(4); 0.39 +/- 0.09 ng/dl) and thyroid stimulating hormone (TSH; 1.12 +/- 0.56 microIU/ml) levels fell within the reptilian range, one order of magnitude lower than mammalian levels. However, cold induced typical mammalian responses: free T(4) levels (0.55 +/- 0.09 ng/dl) and thyroid follicular cell height were significantly greater. Although TSH levels (1.28 +/- 0.83 microIU/ml) were not significantly elevated, thyrotrophs exhibited ultrastructural signs of increased secretory activity. Low thyroid hormone concentrations may contribute substantially to the unusual thermoregulatory mode exhibited by naked mole-rats.

Thyroid Development in Relation to the Development of Endothermy in the Red-Winged Blackbird (Agelaius phoeniceus)

We investigated the development of thyroid function during the transition to endothermy in red-winged blackbirds (Agelaius phoeniceus). Thermoregulatory capabilities of blackbirds improve markedly over their relatively short nestling period (10–12 days), with the most striking improvements occurring between days 6 and 8. We hypothesized that the development of endothermy in these birds is dependent in part on the development of thyroid function. We assessed thyroid development by measuring changes in thyroid gland histology and plasma concentrations of thyroxine (T4) and triiodothyronine (T3) during the nestling period. To gain insight into the role of thyroid maturation in the context of thermoregulation, we compared plasma thyroid hormone profiles in nestlings exposed to cold temperatures to those maintained at thermoneutral temperatures. The overall size of the thyroid (as cross-sectional area) increased during nestling development, with the fastest growth occurring just before the development of endothermy. By day 8, it reached the size typical of that in adults. Follicular cell height of the thyroid glands increased in nestlings up to day 6 and then decreased for the rest of the nestling period. The mean area of individual follicles increased up to day 8 of nestling life and then decreased. Individual nestlings were capable of strong endothermic responses at 7 to 8 days of age and had significantly decreased plasma T4 concentrations following cold exposure, suggesting increased T4 to T3 deiodination to maintain the plasma concentrations of the more metabolically active T3. The patterns of plasma T4 and T3 during nestling development were consistent with those of nestlings of other altricial species of birds that have been studied. Overall, the patterns of thyroid development observed were consistent with the hypothesis that the functional development of the thyroid is critical to the development of endothermic capabilities and that thyroid hormones play a role in endothermic responses to cold temperatures.

Changes in thyroid gland morphology after acute acrylamide exposure.

High exposure to the acrylamide monomer has been associated with neuropathy and neurotoxic effects. Chronic lower exposure causes endocrine disruption associated with thyroid, testicular, and mammary tumors. To investigate mechanisms of endocrine disruption, short-term, low-level oral dosing studies were conducted. Weanling female Fischer 344 rats were acclimatized for two weeks before dosing. Controls were given distilled water by gavage and rats in other groups were given acrylamide at doses of 2 mg/kg/day and 15 mg/kg/day for 2 or 7 days by gavage. Twenty-four h after the last dose, the rats were killed by decapitation. Trunk blood was collected for hormone analyses and tissues for histopathological examination. There were no toxicity-related deaths, no clinical signs of toxicity, and no significant difference in the mean body weight of animal groups. Histopathological examination of select tissues showed no lesions of pathologic significance. Plasma thyroxine (T4), thyroid stimulating hormone (TSH), prolactin (PRL), and pituitary TSH and PRL analyses did not reveal significant changes between control vs. treated rats. In the 7-day study, however, there was a slight dose-dependent increase in plasma T4 and a slight dose-dependent decrease in plasma TSH. Thyroid gland morphometry showed a significant (p < 0.05) decrease in the colloid area and a significant increase (p < 0.05) in the follicular cell height of treated rats as compared to controls. The follicular area shrinkage was similar in both studies. These results show a very early endocrine response to very low levels of toxic insult and opens other venues to further investigate the mechanisms of endocrine disruption by acrylamide.

The Effects of Chlorine Intake on Some Morphometric Parameters of the Thyroid Gland in Lambs

KoAova M., E. Bekeova, M. Levkut: The Effects of Chlorine Intake on Some Morphometric Parameters of the Thyroid Gland in Lambs. Acta Vet. Brno 1999, 68: 191–195. The aim of the study was to determine whether drinking water containing two concentrations of chlorine (0.3 mg . l-1 or 1.8 mg . l-1) affects morphometric indicators of follicular epithelial cells and the diameter of thyroid gland follicles in young lambs. The height of follicular epithelial cells, the diameter of thyroid gland follicles, the frequency of epithelial cell height categories, and the frequency of follicular diameter categories were studied in two groups of lambs after two treatments: one for four weeks and the other for three months. The first group had access to drinking water with 0.3 mg . l -1 chlorine (short-term intake and low Cl concentration) for four weeks. The second group had access to drinking water containing 1.8 mg . l -1 chlorine (long-term intake and high concentration) for three months. A significant increase (P < 0.01) in follicular epithelial cell height and a significant frequency distribution shift to the higher cellular height categories was observed after intake of chlorinated water (0.3 mg . l -1 ), (P < 0.01 for cell height categories 3.1-4 and 4.1-5 µm) for four weeks. There was also a significant shift (P < 0.01 for cell height categories 4.1-5 and 5.1-6 µm) in lambs receiving 1.8 mg . l-1 chlorine in their drinking water for three months as compared to controls. The diameter of thyroid gland follicles significantly decreased (P < 0.01), and the percent of small thyroid follicles significantly increased (for diameter categories 20.1-40 and 40.1-60 µm P < 0.01 and P < 0.05, respectively) in lambs receiving 1.8 mg . l-1 chlorine in drinking water for three months as compared to controls. Our morphometric findings suggest that chlorine interferes with the thyroid metabolism in lambs, however, with no macroscopically detectable changes of the thyroid. Follicular epithelial cell, follicle, cell height, follicular diameter

The effect of fasting and refeeding on thyroid follicule structure and thyroid hormone levels in young and old rats

In order to assess morphological and functional plasticity of the thyroid gland in aging the effects of fasting and refeeding on the thyroid morphology and thyroid hormone serum levels were compared with morphometry and RIA in young and old rats. Young (4 months) and old (24 months) male Wistar rats were fasted for 40 h and sampled, or fasted and fed thereafter for 4 or 24 h. In control (fed) old animals the thyroid follicules were larger, the follicular epithelium was smaller and colloid resorption was smaller than in young rats. ‘Thyroid activation index’ (epithelial volume/colloid volume ratio) was almost twice lower in the thyroids of control old rats. As the result of fasting, height, surface area and volume of epithelial follicular cells decreased in the thyroids of fasted young rats but not in old ones. On the contrary, in thyroids of fasted old rats the dimensions of epithelial cells did not change and thyroid colloid resorption was increased. After 24 h of refeeding, thyroid morphology in both young and old rats did not differ significantly as compared with control animals. Upon fasting, serum levels of thyroxine (T4) and triiodothyronine (T3) decreased by 28 and 38% in young and by 35 and 46% in old rats, respectively. However, T4 and T3 serum concentrations did not differ significantly between age groups in both fed and fasted states. During refeeding the increase in serum thyroxine concentration was delayed in old rats as compared with young ones. The results of morphological, morphometric and hormonal investigations indicate the existence of age-related changes in the structure and function of thyroid follicular cells.

MORPHOMETRY CHANCES IN THE PREPUBERTAL FEMALE RAT THYROID GLAND FOLLOWING ACUTE EXPOSURE TO 2,2′,4,4′-TETRACHLOROBIPHENYL AND AROCLOR 1242

Weanling female Sprague-Dawley rats were given either 2 or 5 consecutive daily doses of 30 mg/kg 2,2′,4,4′-tetrachlorobiphenyl (CB 47) or a total dose of 120 mg/kg Aroclor 1242 divided into 2, 3, or 5 daily doses by intraperitoneal injection. One day after the final dose, serum total thyroxine (T4) was determined and thyroid glands were collected and prepared for morphometry analysis. Serum T4 increased between 20 and 25 d of age, but declined to 35–52% of controls by d 25 in PCB-treated rats. In rats receiving only 2 doses of CB 47, the declines in serum T4 were more modest but the thyroid follicular epithelial cell height increased from 9 µm to 10–12 µm and the colloid area decreased from 1100 µm2 to 800–900 µm2. In Aroclor 1242-treated rats, follicular cell height increases and colloid area decreases were somewhat greater; serum T4 was higher (partially restored) in the rats having received earlier doses. The rapid response of the thyroid gland to moderate decreases in serum T4 attenuates the observed decrease in T4 and may mask effects on T4 metabolism in short-term structure-activity studies. Morphometric measurements may be helpful in characterizing early and/or transient responses to toxicants such as PCBs that have multiple endocrine disrupting effects.