Fig. 10

Fig. 10 Disruption of hormonal and paracrine signaling during masculinization in triploid zebrafish. (A). Diferentially expressed genes in triploid (3n) zebrafish gonads compared to the diploid controls (2n) based on RNAseq. Differentially expressed genes were Kelch-like protein 10 (klhl10), dehydrogenase/reductase member 3a (dhrs3a), cytochrome P450 family 26 subfamily B member 1 (cyp26b1), prostaglandin E receptor 2a (ptger2a), deodeinase 1 (dio1), follicle-stimulating hormone receptor (fshr), insulin-like growth factor 3 (igf3), cytochrome P450 family 11 subfamily A member 1 (cyp11a1), steroidogeneic acute regulatory protein (star) and antiMullerian hormone (amh) and Dishevelled-binding antagonist of beta-catenin 1 (dact1). Asterisks denote significant differences between treatments. Normalized counts and statistical significance were obtained using DESeq2 median of ratios normalization and negative binomial differential expression modeling. (B). Constructed gene regulatory network in the gonads of triploid (3n) zebrafish at 120 days post-fertilization compared with diploid (2n) controls. All triploids developed as males. The figure highlights key molecular pathways driving masculinization in triploid zebrafish. Suppression of fshr and amh released igf3 from inhibition, stimulating spermatogenesis directly and potentially through sustained β-catenin activation (facilitated by dact1 downregulation). Reduced ptger2a and dio1 expression, together with enhanced retinoic acid degradation by cyp26b1 and dhrs3a, further promoted spermatogonial proliferation while impairing meiotic progression and repressing ovarian differentiation. Finally, klhl10 downregulation provides a molecular explanation for the reduced fertility consistently observed in male triploids. Upward/downward arrows indicate up- or downregulation of the adjacent gene; colored solid lines represent upregulation, colored dashed lines downregulation, and grey lines genes not differentially expressed but active in baseline gonadal physiology. Line endings denote stimulatory or inhibitory interactions. Abbreviations: spermatogonia An undifferentiated (Spg Aund), spermatogonia A differentiated (SpgAdiff), spermatogonia B (SpgB), spermatocyte (Spc), spermatid (Spt), spermatozoa (Stz). Created in BioRender. MARTINEZ, A. (2025) https://BioRender.com/q3sjavl.