Protocol for culture of zebrafishdeveloping ovaries as an ex vivo system for investigating early oogenesis and ovarian development.

A fundamental understanding of early oogenesis is lacking, and zebrafish serve as an excellent model for investigation, based largely on in vivo approaches. We present a protocol for long-term zebrafish ovary culture that supports physiological early oogenesis ex vivo, enabling new experimental approaches. We outline procedures for culture plate and media preparation, ovary dissection, embedding-media-free mounting, and culture handling. In addition, we describe workflows for ovary evaluation and rapid functional assays. For complete details on the use and execution of this protocol, please refer to Shawahny et al.1.