Abstract
BackgroundSex-related genes play a crucial role in gonadal differentiation into testes or ovaries. However, the genetic control of gonadal differentiation in Muscovy ducks remains unknown. Therefore, the objective of our study was to screen new candidate genes associated with ovarian and testicular development.ResultsIn this study, 24 males before gonadal differentiation (MB), 24 females before gonadal differentiation (FB), 24 males after gonadal differentiation (MA) and 24 females after gonadal differentiation (FA) were selected from Putian Muscovy ducks, forming 4 groups. RNA-Seq revealed 101.76 Gb of clean reads and 2800 differentially expressed genes (DEGs), including 46 in MB vs FB, 609 in MA vs FA, 1027 in FA vs FB, and 1118 in MA vs MB. A total of 146 signalling pathways were enriched by KEGG analysis, among which 20, 108, 108 and 116 signalling pathways were obtained in MB vs FB, MA vs MB, MA vs FA and FA vs FB, respectively. In further GO and KEGG analyses, a total of 21 candidate genes related to gonad differentiation and development in Muscovy ducks were screened. Among these, 9 genes were involved in the differentiation and development of the testes, and 12 genes were involved in the differentiation and development of the ovaries. In addition, RNA-Seq data revealed 2744 novel genes.ConclusionsRNA-Seq data revealed 21 genes related to gonadal differentiation and development in Muscovy ducks. We further identified 12 genes, namely, WNT5B, HTRA3, RSPO3, BMP3, HNRNPK, NIPBL, CREB3L4, DKK3, UBE2R2, UBPL3KCMF1, ANXA2, and OSR1, involved in the differentiation and development of ovaries. Moreover, 9 genes, namely, TTN, ATP5A1, DMRT1, DMRT3, AMH, MAP3K1, PIK3R1, AGT and ADAMTSL1, were related to the differentiation and development of testes. Moreover, after gonadal differentiation, DMRT3, AMH, PIK3R1, ADAMTSL1, AGT and TTN were specifically highly expressed in males. WNT5B, ANXA2 and OSR1 were specifically highly expressed in females. These results provide valuable information for studies on the sex control of Muscovy ducks and reveal novel candidate genes for the differentiation and development of testes and ovaries.
Highlights
Sex-related genes play a crucial role in gonadal differentiation into testes or ovaries
Overview of RNA-Seq results In order to identify gonadal differentiation genes in Muscovy ducks, transcriptome analysis of Muscovy duck gonads was performed on males before gonadal differentiation (MB), females before gonadal differentiation (FB), males after gonadal differentiation (MA), and females after gonadal differentiation (FA)
Anti-Mullerian hormone (AMH) is significantly upregulated in MA vs FA and MA vs MB, that is, AMH is expressed in males after gonadal differentiation, so it is a candidate gene for the testis in Muscovy ducks
Summary
Sex-related genes play a crucial role in gonadal differentiation into testes or ovaries. Sex in birds and mammals is governed by sex chromosomes [1], and sex determination genes on sex chromosomes initiate sex differentiation and regulate the differentiation of gonads into ovaries or testes [2]. Screening for novel regulators of male and female embryos is highly necessary to enhance our understanding of gonadal differentiation. In ducks and other avian species, there is a female heterogametic system of ZZ/ZW sex chromosomes [6], and their sex may be governed by genes on the W chromosome [7]. Studies have found that the mechanism of sex differentiation in birds is different from that in mammals [6, 8]. The exact role of avian sexdetermining genes on Z and W has yet to be completely resolved
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