Menopause is associated with an accumulation of visceral fat. An emerging concept suggests that relatively elevated levels of circulating androgens compared with estrogens in post-menopausal women underlie this shift in body-fat distribution. In this study we administered dihydrotestosterone (DHT) to ovariectomised (ovx) mice to examine the effect of relative androgen excess on adipose tissue distribution and function in estrogen-deficient mice. Compared with controls, DHT-treated mice exhibited increased body weight and visceral fat mass associated with triglyceride accumulation. Phosphorylation of AMP-activated-protein-kinase (AMPK) and Acetyl-CoA-Carboxylase (ACC) was significantly decreased by DHT in visceral fat. In 3T3-L1 cells, DHT decreased phosphorylation of AMPK in a dose-dependent manner. In addition, DHT increased the expression of lipogenic genes (Fatty-Acid-Synthase, Sterol Regulatory Element Binding Protein-2 and Lipoprotein Lipase) in visceral fat. These data provide the first in vivo evidence that an increased androgen-estrogen ratio can promote visceral fat accumulation by inhibiting AMPK activation and stimulating lipogenesis.