title: A Tandem Duplicate of Anti-Müllerian Hormone with a Missense SNP on the Y Chromosome Is Essential for Male Sex Determination in Nile Tilapia, Oreochromis niloticus.
abstract: Variation in the TGF-β signaling pathway is emerging as an important mechanism by which gonadal sex determination is controlled in teleosts. Here we show that amhy, a Y-specific duplicate of the anti-Müllerian hormone (amh) gene, induces male sex determination in Nile tilapia. amhy is a tandem duplicate located immediately downstream of amhΔ-y on the Y chromosome. The coding sequence of amhy was identical to the X-linked amh (amh) except a missense SNP (C/T) which changes an amino acid (Ser/Leu92) in the N-terminal region. amhy lacks 5608 bp of promoter sequence that is found in the X-linked amh homolog. The amhΔ-y contains several insertions and deletions in the promoter region, and even a 5 bp insertion in exonVI that results in a premature stop codon and thus a truncated protein product lacking the TGF-β binding domain. Both amhy and amhΔ-y expression is restricted to XY gonads from 5 days after hatching (dah) onwards. CRISPR/Cas9 knockout of amhy in XY fish resulted in male to female sex reversal, while mutation of amhΔ-y alone could not. In contrast, overexpression of Amhy in XX fish, using a fosmid transgene that carries the amhy/amhΔ-y haplotype or a vector containing amhy ORF under the control of CMV promoter, resulted in female to male sex reversal, while overexpression of AmhΔ-y alone in XX fish could not. Knockout of the anti-Müllerian hormone receptor type II (amhrII) in XY fish also resulted in 100% complete male to female sex reversal. Taken together, these results strongly suggest that the duplicated amhy with a missense SNP is the candidate sex determining gene and amhy/amhrII signal is essential for male sex determination in Nile tilapia. These findings highlight the conserved roles of TGF-β signaling pathway in fish sex determination.
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Minghui
authors.initials: M
authors.lastname: Li
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Yunlv
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Jiue
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Hongjuan
authors.initials: H
authors.lastname: Shi
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Zeng
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Ye
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Dongneng
authors.initials: D
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Linyan
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Sun
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Tao
authors.affiliation: Solution-Oriented Research for Science and Technology (SORST), Laboratory of Reproductive Biology, National Institute for Basic Biology, Okazaki, Japan; South Ehime Fisheries Research Center, Ehime University, Matsuyama, Japan.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Nagahama
authors.affiliation: Department of Biology, University of Maryland, College Park, Maryland, United States of America.
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Kocher
authors.affiliation: Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Science of Chongqing, School of Life Science, Southwest University, Chongqing, China.
authors.forename: Deshou
authors.initials: D
authors.lastname: Wang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D054304:Anti-Mullerian Hormone / Q000235:genetics*; D005260:Female; D020440:Gene Duplication; D008040:Genetic Linkage; D008297:Male; D020125:Mutation, Missense; D020641:Polymorphism, Single Nucleotide; D019849:Sex Determination Processes; D017210:Tilapia / Q000235:genetics*; D014998:Y Chromosome
chemical_list: D054304:Anti-Mullerian Hormone