Abstract: Gonadal soma-derived factor（Gsdf） is a member of the transforming growth factor-β（TGF-β） superfamily and acts as a male initiation factor in medaka（Oryzias latipes）, however, the targeted genes under the regulation of Gsdf is still unclear. We performed RNA-sequence and RT-qPCR analysis to identify the downstream gene expression responsible for Gsdf signaling. According to the comparison of RNA-sequence profiles among normal and gsdf deficient adult gonads, the expression of zc4 h2（zinc finger C4 H2-type containing） was significantly different(q＜0.05 and ■ between the gsdf-null XY ovary and normal XX ovary. ZC4 H2 is involved in BMP/SMADs-mediated neural progenitor development through the physical binding of SMAD3, and one of the pathogenic factors of human Wieacker-Wolff syndrome linked to the X chromosome. In medaka, zc4 h2 is expressed in multiple tissues and enriched in brain and gonad, whereas the expression level in male is higher than that in female. Compared with normal XX ovaries, the transcripts of zc4 h2 3′ untranslated region（3′ UTR） in gsdf deficient XY ovaries significantly increased, suggesting that Gsdf-Smad3 signaling may affect the level and the stability of zc4 h2 transcripts in gonad development. The amino acid sequence and functional domains of Gsdf-Zc4 h2-Smad3 are identical among species, indicating that there is a conserved and universal mechanism of sexual differentiation in vertebrate brain and gonad development, thus these results provide insights into the molecular mechanism for the differentiation and development of vertebrate germ stem cells and neural stem cells.