Zebrafish sex determination, gonad differentiation and reproduction are far from being fully understood. Although the mode of sex determination is still being disputed, most experimental data point towards the lack of sex chromosomes and a multigenic sex determination system. Secondary effects from the environment and/or (xeno)hormones may influence the process, resulting in biased sex ratios. The exact time point of sex determination is unknown. Gonad differentiation involves a compulsory 'juvenile ovary' stage with subsequent transformation of the gonad into a testis in males. As the latter is a late event, there is a delay between sex determination and testis differentiation in zebrafish, in contrast to mammals. Information on the expression of several candidate genes thought to be involved in these processes has been supplemented with data from large-scale gonadal transcriptomic studies. New approaches and methodologies provide hope that answers to a number of important questions will be deciphered in the future.