Yang, Z.; Luo, T.; Zhang, Y.; Sun, Y.

The ricefield eel (Monopterus albus), an economically important aquaculture species in China, is the only freshwater teleost fish that exhibits protogynous hermaphroditism. While progress has been made towards understanding the sex determination and differentiation of this species, the understanding mechanisms remain elusive. Here we show that warm temperature promotes gonadal transformation by up-regulating testicular differentiation genes such as dmrt1/sox9a in ovaries. Trpv4, a Ca2+-permeable cation channel expressed in gonads, is highly sensitive to ambient temperature and mediates warm temperature-driven sex change of ricefield eel. In female fish reared at cool temperature, injection of Trpv4 agonist into the ovaries leads to significant up-regulation of testicular differentiation genes, and in female fish exposed to warm temperature, Trpv4 antagonist administration or trpv4 siRNA knockdown inhibits temperature-induced male gene expression. pStat3 signaling is downstream of Trpv4 and transduces Trpv4-controlled calcium signaling into the sex determination cascades. Inhibition of pStat3 activity prevents the up-regulation of male genes by warm temperature treatment and ovarian injection of Trpv4 agonist, whereas activation of pStat3 is sufficient to induce the expression of male genes, even in the presence of Trpv4 antagonist. pStat3 binds and activate jmjd3/kdm6b, an activator of the male gene dmrt1. Consistently, ovarian injection of Kdm6b inhibitor blocks the up-regulation of male genes by warm temperature treatment. We propose that environmental factors such as warm temperature promote gonadal transformation of ricefield eel via the Trpv4-pStat3-Kdm6b axis. Our results provide new insights into the molecular mechanism underlying natural sex change of ricefield eel, which will be useful for sex control in aquaculture.