Abstract

Gonadal differentiation is tightly regulated by the initial sex determining gene and the downstream sex-related genes in vertebrates. However, sex change in fish can alter the sexual fate from one sex to the other. Chemical-induced maleness in the protogynous orange-spotted grouper is transient, and a reversible sex change occurs after the chemical treatment is withdrawn. We used these characteristics to study Amh signaling during bi-directional sex change in the grouper. We successfully induced the female-to-male sex change by chemical (aromatase inhibitor, AI, or methyltestosterone, MT) treatment. A dormant gonad (a low proliferation rate of early germ cells and no characteristics of both sexes) was found during the transient phase of reversible male-to-female sex change after the withdrawal of chemical administration. Our results showed that amh (anti-mullerian hormone) and its receptor amhr2 (anti-mullerian hormone receptor type 2) were significantly increased in the gonads during the process of female-to-male sex change. Amh is expressed in the Sertoli cells surrounding the type A spermatogonia in the female-to-male grouper. Male-related gene (dmrt1 and sox9) expression was immediately decreased in MT-terminated males during the reversible male-to-female sex change. However, Amh expression was found in the surrounding cells of type A spermatogonia-like cells during the transient phase of reversible male-to-female sex change. This phenomenon is correlated with the dormancy of type A spermatogonia-like cells. Thus, Amh signaling is suggested to play roles in regulating male differentiation during the female-to-male sex change and in inhibiting type-A spermatogonia-like cell proliferation/differentiation during the reversible male-to-female sex change. We suggest that Amh signaling might play dual roles during bi-directional sex change in grouper.

Highlights

  • In mammals, sex determination results from the initial switch of either the Sry-dependent, testis-differentiating or Sry-independent, ovary-differentiating molecular cascade in an exclusive manner [1]

  • Our results indicate that Amh signaling might regulate male differentiation during female-to-male sex change and prevent advanced differentiation in early germ cells during amh/Amh expression patterns during bi-directional sex change in protogynous grouper reversible male-to-female sex change

  • Amh antibody can’t recognize the N-terminal and C-terminal Amh fragment after Amh processing through the potential plasmin protease cleavage site (RGLRATR). These results reveal that plasmin protease cleavage site (RGLRATR, amino acid 408–414 in grouper) upstream of the Tgfb-domain that may be necessary for the processing of the grouper Amh

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Summary

Introduction

In mammals (mice), sex determination results from the initial switch of either the Sry-dependent, testis-differentiating or Sry-independent, ovary-differentiating molecular cascade in an exclusive manner [1]. October 10, 2017 amh/Amh expression patterns during bi-directional sex change in protogynous grouper in the development of an ovotestis [2]. These ovotestes can be caused by an insufficient or delayed expression of the testis-related gene in the XY gonad [3,4,5] or loss of the ovary-related gene in the XX gonad [6]. Gonadal differentiation is tightly regulated by the initial sexdetermining gene and downstream sex-related genes in mammals. The key gene to switch controlling the gonadal sex differentiation cascade leading to the development of one sex but the regression of the other sex remains unclear in hermaphroditic fish

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