Abstract.To examine the effects of diethylstilbestrol (DES) on male pubertal development and thyroid function, juvenile male Sprague-Dawley rats were given DES daily by oral intubation at doses of 10, 20 and 40 μg/kg/day from postnatal day 33 for 20 days. Prepuce separation was significantly delayed at the dose of 20 μg/kg/day and above in the DES-treated rats. DES treatment induced a significant reduction in the weights of testes, epididymides, the ventral prostate, seminal vesicles plus coagulating glands and fluid, levator ani bulbocavernosus muscles, Cowper's glands and the glans penis. The weights of the liver and adrenals increased in the DES-treated animals. DES caused a dosedependent reduction in germ cells; in particular the spermatids were mainly affected. The serum levels of testosterone and luteinizing hormone were significantly reduced in the DES-treated groups, but that of estradiol decreased. No differences were observed in the serum thyroxine levels of the control and DES-treated groups. In microscopic observation of the DES-treated animals, degeneration of germ cells and tubular atrophy in the testis were noted, but there were no microscopic changes in the thyroid. These results indicate that DES affected the pubertal development of juvenile male rats and that its mode of action may be related to alterations in hormone levels. Key words: Diethylstilbestrol, Male rat, Pubertal development (J. Reprod. Dev. 55: [461][462][463][464][465][466] 2009) oncerns have been raised about the potential adverse effects of environment chemicals that mimic the actions of estrogen on the reproduction and development of humans and wildlife species. Exposure of the developing male to exogenous estrogenic compounds can result in reproductive and developmental abnormalities and an increase in human male reproductive disorders, which include testicular cancer, cryptorchidism, hypospadias and low sperm counts [1][2][3][4]. Neonatal treatment with estrogens causes alterations in germ cell development and permanent impairment of testicular function [5][6][7][8][9]. As perinatal exposure of male animals and humans to synthetic estrogens can induce similar adverse changes in male reproductive health, it has been hypothesized that inappropriate exposure to estrogens during juvenile periods may interfere with the development of the reproductive system and with the sperm count in the adult [10,11]. Faced with growing concern that environmental chemicals might impair human and animal fertility, it is important to investigate the possible influences of these substances on mammalian sexual differentiation and genital development [12,13].Diethylstilbestrol (DES) is a stilbene, a non-steroid estrogen, that can bind to estrogen receptors (ERs) and is widely used as a model estrogen for the study of estrogenic effects on the male reproductive function [13][14][15][16]. DES has been observed to have endocrine and reproductive toxicity in rats and mice. It is active over a wide dose range, showing effects on the mouse ventral prost...