Amh/Amhr2 Signaling Causes Masculinization by Inhibiting Estrogen Synthesis during Gonadal Sex Differentiation in Japanese Flounder (Paralichthys olivaceus)
Abstract
:1. Introduction
2. Results
2.1. Expression Analysis of amh and amhr2 in Gonads in XY and XX Flounder
2.2. Generation of amhr2-Deficient Mutant Using CRISPR/Cas9 System in Japanese Flounder
2.3. amhr2 Mutants Showed Phenotypic Sex Reversal from Male to Female
2.4. Effect of Fadrozole Treatment on Gonadal Phenotype of amhr2 Mutants
3. Discussion
4. Materials and Methods
4.1. Experimental Animals
4.2. Determination of Genetic Sex
4.3. Expression Analysis by Real-Time PCR
4.4. Histological Analysis of amhr2 Transcripts Using In Situ Hybridization
4.5. Generation of amhr2 Mutant Using CRISPR/Cas9 System
4.6. Analysis of Genotype in amhr2 Mutants
4.7. Histological Observations
4.8. Experimental Treatment with Fadrozole
4.9. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Types of Mutations | ||||||
---|---|---|---|---|---|---|
Genetic Sex | Phenotypic Sex (Gonad) | Number of Clones Sequenced | Wide Deletion | Short Deletion | No Mutation | |
wild-type | XY | ♂ (Testis) | 4 | 0 | 0 | 4 |
XX | ♀ (Ovary) | 4 | 0 | 0 | 4 | |
amhr2 mutant | XY | ♀ (Ovary) | 16 | 15 | 1 | 0 |
XY | ♀ (Ovary) | 16 | 13 | 3 | 0 | |
XY | ♀ (Ovary) | 16 | 15 | 1 | 0 | |
XX | ♀ (Ovary) | 8 | 8 | 0 | 0 | |
XX | ♀ (Ovary) | 8 | 6 | 2 | 0 |
Treatment | Genotype | Genetic Sex | ♂ (Testis) | ♀ (Ovary) |
---|---|---|---|---|
Control | wild-type | XY | 9 | 0 |
Control | wild-type | XX | 0 | 11 |
Control | amhr2 mutant | XY | 0 | 6 |
Control | amhr2 mutant | XX | 0 | 4 |
Fadrozole | amhr2 mutant | XY | 5 | 0 |
Fadrozole | amhr2 mutant | XX | 5 | 0 |
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Yamaguchi, T.; Kitano, T. Amh/Amhr2 Signaling Causes Masculinization by Inhibiting Estrogen Synthesis during Gonadal Sex Differentiation in Japanese Flounder (Paralichthys olivaceus). Int. J. Mol. Sci. 2023, 24, 2480. https://doi.org/10.3390/ijms24032480
Yamaguchi T, Kitano T. Amh/Amhr2 Signaling Causes Masculinization by Inhibiting Estrogen Synthesis during Gonadal Sex Differentiation in Japanese Flounder (Paralichthys olivaceus). International Journal of Molecular Sciences. 2023; 24(3):2480. https://doi.org/10.3390/ijms24032480
Chicago/Turabian StyleYamaguchi, Toshiya, and Takeshi Kitano. 2023. "Amh/Amhr2 Signaling Causes Masculinization by Inhibiting Estrogen Synthesis during Gonadal Sex Differentiation in Japanese Flounder (Paralichthys olivaceus)" International Journal of Molecular Sciences 24, no. 3: 2480. https://doi.org/10.3390/ijms24032480