Ability of the Hershberger assay protocol to detect thyroid function modulators.
In vivo screening methods for detection of thyroid function modulators are now under development in many research laboratories. We assessed the applicability of the Hershberger assay protocol to screen for thyroid function modulators. In experiment 1, castrated male BrlHan WIST@Jcl (GALAS) rats were administered a potent thyroid peroxidase inhibitor, 3-amino-1,2,4-triazole (AT), in doses of 0, 40, 200, and 1,000 mg/kg/day with gravimetric endpoint, and in experiment 2, castrated and intact male rats were administered in doses of 0, 40, and 200 mg/kg/day, with quantification of the extent of hypertrophy of the thyroid epithelium, to assess the effects of castration, by gavage to 8-week-old for 10 consecutive days. At necropsy of both experiments, the thyroid glands and hypophysis were collected and fixed with 10% neutral-buffered formalin. To avoid crushing during weighing because of their fragility, the thyroid glands and hypophysis were weighed approximately 24 h after fixation with 10% neutral-buffered formalin. All animals were sacrificed approximately 24 h after the final dose. In experiment 2, the thyroid glands of all animals were stained with hematoxylin and eosin for histological examination and morphometry of follicular epithelial height. In experiment 1, absolute and relative thyroid weights in all of the AT groups were statistically increased in a dose-dependent manner, regardless of the testosterone propionate (TP)-injection. In experiment 2, the results showed a significant increase in thyroid weight in the 200 mg/kg groups of both castrated and intact rats. Hypophyseal weight was unaltered by AT, but comparison of vehicle-treated groups showed that the hypophyseal weight of the castrated rats was greater than that of the intact rats. Enlarged thyroid glands were observed in the AT-treated rats at necropsy. Histological examination of the thyroid glands of all the AT-treated animals showed hypertrophy and hyperplasia of the follicular epithelial cells, and the height of follicular epithelium of the thyroid glands increased in a dose-dependent manner in both the castrated and intact rats. In experiment 1, assessment of the (anti-) androgenic action of AT in seminal vesicle weight revealed a significant increase in the 200 and 1,000 mg/kg + TP groups in a dose-dependent manner. These results suggest that the effect of AT can be detected by the Hershberger assay 10-day administration protocol and may be useful for screening for thyroid function modulators regardless of whether the animals have been castrated.
Noda
Shuji
S
Chemicals Assessment Center, Chemicals Evaluation and Research Institute, 822, 3-chome, Ishii-machi, Hita-shi, Oita 877-0061, Japan. noda-shuji@ceri.jp
Muroi
Takako
T
Takakura
Saori
S
Sakamoto
Satoko
S
Takatsuki
Mineo
M
Yamasaki
Kanji
K
Tateyama
Susumu
S
Yamaguchi
Ryoji
R
D000640:Amitrole / Q000633:toxicity*; D000818:Animals; D001681:Biological Assay; D002369:Castration; D004791:Enzyme Inhibitors / Q000633:toxicity; D006540:Herbicides / Q000633:toxicity; D008297:Male; D009929:Organ Size / Q000187:drug effects; D009195:Peroxidase / Q000037:antagonists & inhibitors; D051381:Rats; D011919:Rats, Inbred Strains; D012669:Seminal Vesicles / Q000187:drug effects / Q000473:pathology; D013961:Thyroid Gland / Q000187:drug effects* / Q000473:pathology
D004791:Enzyme Inhibitors; D006540:Herbicides; D009195:Peroxidase; D000640:Amitrole