We examined the effects of dexamethasone (DEX) on the expressions of key proteins concerned with energy metabolism in brown adipocytes during their differentiation in primary culture. Transcripts of the uncoupling protein (UCP), lipoprotein lipase (LPL) and CCAAT enhancer binding protein alpha (C/EBP alpha) genes were observed in brown adipocytes cultured in the presence of insulin and thyroid hormones but in the absence of DEX. However, the mRNA level of UCP decreased with the culture period after confluence, and significant mRNA encoding type-1 glucose transporter (GLUT1) was detected in brown adipocytes cultured without DEX, whereas type-4 glucose transporter (GLUT4) was predominantly expressed in mature brown adipocytes in vivo. In contrast, DEX added after confluence consistently elevated the mRNA levels of UCP, LPL and C/EBP alpha, and repressed the level of GLUT1 in a manner synchronized with increase in the level of GLUT4. Therefore, it is concluded that DEX as well as insulin and thyroid hormones is essential for differentiation of brown adipose precursor cells into mature cells that are similar to brown adipocytes in vivo.