During development all zebrafish males first develop a “juvenile ovary” that later degenerates and transforms into a testis. In this study, individuals undergoing gonadal transformation were identified from a vas::egfp transgenic line and used for gene expression analysis of anti-Müllerian hormone (amh), ovarian aromatase (cyp19a1a) and 11β-hydroxylase (cyp11b, also known as P45011β) by real-time polymerase chain reaction and in situ hybridization. In the “normal (i.e., nontransforming) juvenile ovary” cyp19a1a was expressed around the oocytes, but cyp11b and amh could not be detected. During gonadal transformation cyp19a1a was down-regulated and could not be detected anymore; in contrast amh was up-regulated and highly expressed at similar regions where cyp19a1a had been expressed earlier. Furthermore, the normalized transcript levels of cyp19a1a and amh showed a reciprocal picture, i.e., the higher was the level of amh, the lower was that of cyp19a1a. Expression of cyp11b was also up-regulated but later than amh, and its localization was not related to the position of degenerating oocytes. These data indicate that amh is a candidate gene down-regulating cyp19a1a, leading to “juvenile ovary-to-testis” transformation. Whereas, cyp11b or its product, 11-ketotestosterone, is unlikely to be the inducer of zebrafish gonad transformation, as proposed earlier for some protogynous hermaphroditic fish species. Developmental Dynamics 236:1329–1338, 2007. © 2007 Wiley-Liss, Inc.