Abstract
Zinc finger AN1-type domain 3 (ZFAND3) is essential for spermatogenesis in mice. However, its function in teleosts remains unclear. In this study, we characterized the ZFAND3 gene (termed as OsZFAND3) in an important food fish, tilapia. The OsZFAND3 cDNA sequence is 1,050 bp in length, containing an ORF of 615 bp, which encodes a putative peptide of 204 amino acid residues. Quantitative real-time PCR revealed that the OsZFAND3 transcripts were exclusively expressed in the testis and ovary. In situ hybridization showed that the high expression of OsZFAND3 transcripts was predominantly localized in the spermatocyte and spermatid. These results suggest that OsZFAND3 is involved in male germ cell maturation. Three single nucleotide polymorphisms (SNPs) were detected in the introns of OsZFAND3. The OsZFAND3 gene was mapped in the sex-determining locus on linkage group 1 (LG1). The three SNPs in the OsZFAND3 gene were strictly associated with sex phenotype, suggesting that the OsZFAND3 gene is tightly linked to the sex-determining locus. Our study provides new insights into the functions of the OsZFAND3 gene in tilapia and a foundation for further detailed analysis of the OsZFAND3 gene in sex determination and differentiation.
Highlights
Tilapia is the common name for a group of cichlid fishes native to both fresh and salt water in Africa and the Middle East
Sex-determining loci were mapped to LG1 and LG23 in Nile tilapia (Oreochromis niloticus)[17,18], LG3 in hybrid population of O. niloticus × O. aureus[19], and LG1 and LG22 in Mozambique tilapia and red tilapia[13]
SNP1 and SNP2 were located in the first intron, while SNP3 was found in the fifth intron
Summary
The best way to obtain a male monosex population is through genetic control by developing YY “super males” which can sire more male progenies[15] This technology is reliable and consistent, and more and more sex-associated markers were identified and utilized in various tilapiine species. By using RAD sequencing, a set of SNP markers highly associated with phenotypic sex were identified on LG1 in the O. niloticus families[20]. Our previous study showed that one sex-determining locus was mapped onto LG1 in a tilapia family[13], no potential sex-determining gene has been identified on LG1 in this family yet. The purpose of this study was to characterize the OsZFAND3 gene, which was mapped onto the sex-determining locus on LG1 in the family of hybrid tilapia (Oreochromis spp.) constructed by crossing a Mozambique tilapia male and a red tilapia female. Our results suggest that OsZFAND3 is tightly linked to the sex-determining locus, and it may play an essential role in male germ cell maturation
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