Zebrafish (Danio rerio) studies describe before the onset of mature gonads differentiation all individuals go through a "juvenile ovary" stage. However, the sequential events of the early zebrafish gonad differentiation are still not described in full detail and recent works indicate that some individuals never form a "juvenile ovary" structure. Therefore, the present study aimed to confirm the existence of two processes of zebrafish male differentiation. For this purpose, every two days between 20 and 30 days post-fertilization (dpf) zebrafish were collected for a stereological analysis of the differentiating gonads. The histological evaluation showed that prior to 22 dpf, zebrafish gonads were still undifferentiated. At 24 dpf, some individuals started to present a "juvenile ovary" and from 26 to 30 dpf, it was possible to discern two processes of gonad development. The majority of the individuals (80 %) developed a "juvenile ovary", while in the remaining (20 %) it was not possible to detect this structure. The results of the present study show the existence of two distinct processes of zebrafish male gonad development, indicating that not all individuals go through the "juvenile ovary" stage.
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