A total of 160 Sprague-Dawley rats averaging in body weight, were used for histopathological and biochemical studies of toxicity of bis (tri-n-butyltin) oxide (TBTO). Short-term effects and long-term effects were examined. LD50 of TBTO by gastric tubing was 197 mg/kg. The effects of a single dose of TBTO were transient, and rapid recovery followed. TBTO at a a single dose of 100 mg/kg, or a total dose of 390 to 780 mg/kg in 13 weeks, or 780 to 1,560 mg/kg in 26 weeks, caused adrenal hypertrophy, flattening of the thyroid epithelium and atrophy of the thymus and lymph nodes. A single or repeated administration induced swelling and vacuolation of aldehyde-fuchsin-positive cells in the adenohypophysis. The immunohistochemical stainability of the cytoplasm of ACTH-cells was markedly depressed 24 hours after TBTO treatment, and that of the cytoplasm of TSH-cells was enhanced. Analogous an inverse relationship was also manifested in the hormonal levels of serum: levels of cortisol were elevated, while those of T-4 and TSH were depressed.