Zebrafish has been used increasingly as a good animal model for a number of studies. To facilitate the use of this zebrafish model, the current report put emphasis on the study of two steroidogenic genes: cyp11a1 and hsd3b. These two genes encode enzymes that catalyze the first two steps of the steroidogenic pathway, and both enzymes are important for the synthesis of all steroids. Zebrafish cyp11a1 and hsd3b genes are expressed in the same cells in the gonads and interrenal gland. The interrenal gland is the counterpart of mammalian adrenal; it is located inside the head kidney and is developed parallel to the development of the pronephros. In addition, cyp11a1 and hsd3b are also expressed in the blastomeres of the early embryos before gastrulation, and in the extra-embryonic yolk syncytial layer during gastrulation. This early expression implies a novel role of steroids at gastrulation.