Abstract Introduction: This work describes various aspects of early gonadal development of female and male in pre-metamorphic tadpoles (Lithobates catesbeianus) at Gosner stage 25. Materials and Methods: Ovaries and testicles were prepared for routine light microscopy for morphological study and for acridine orange technique fluroescent microscopy for observation of RNA cytoplasm activity. Results: The results showed that female gonads at Gosner stage 25 predominated primary and secondary oogonias, as well as primary, secondary and tertiary oocytes. The developing testicle presented primary spermatogonia (I or A) and secondary spermatogonia (II or B), and as well as primary and secondary spermatocyte. All this cell phases were morphologically characterized and the cell sizes measured. In pre-metamorphic testes the somniferous duct are not developed and the vasa deferentia is opened. Conclusion: At this point, it was possible differentiate ovary from testes does not for the gonadal cells, but for the general anatomy of the organs, being the ovary a spheroid structure and the testicle an elongated tubule.