Comparative Transcriptome Analysis of Three Types of Gonads in Northern Snakehead (Channa argus)
HUANG Su-Jing1,2, WU Yu-Xia1,2, XIA Wei-Wei1,2, ZHANG Xiao-Tian1,2, ZHAO Jian1,2, LI Kai-Bin1, LUO Qing1, LIU Hai-Yang1, FEI Shu-Zhan1, CHEN Kun-Ci1,2, OU Mi1,*
1 Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences/Key Laboratory of Utilization and Aquaculture of Tropical and Subtropical Aquatic Resources, Ministry of Agriculture and Rural Affairs, Guangzhou 510380, China; 2 College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China
Abstract：Northern snakehead (Channa argus) is an important freshwater fish in China with significantly sexual dimorphism. In order to illuminate the molecular mechanism of sex differentiation and gonadal development of C. argus, and explore the functional genes and signaling pathways that differentially express in testis and ovary, RNA-seq was performed on gonads of 6-month-old normal XX females (XX-F), normal XY males (XY-M), and YY super-males (YY-M) by Illumina HiSeq 2000. Correlation analysis of pairwise samples of gonadal tissues indicated that similar expression patterns were found in XY-M and YY-M testis, which were different from that in XX-F ovary, and the result was consistent with the histological observation of gonads. A tremendous amounts of differentially expressed genes (DEGs) were found in the pairwise comparison among XX-F, XY-M, and YY-M gonads. 12 317 DEGs were obtained in XY-M testis compared with XX-F ovary, in which 5 427 were up-regulated and 6 890 were down-regulated. 212 DEGs were obtained in XY-M testis compared with YY-M testis, in which 59 were up-regulated and 153 were down-regulated. 11 282 DEGs were obtained in YY-M testis compared with XX-F ovary, in which 5 038 were up-regulated and 6 244 were down-regulated. qRT-PCR was used to verify DEGs that were related to sex differentiation and gonadal development with significant differences among testis and ovary. It displayed that 10 genes had relatively high expression in testis of XY-M and YY-M, such as anti-müllerian hormone receptor type 2 (Amhr2), anti-müllerian hormone (Amh), doublesex-mab3-related transcription factor1 (Dmrt1) and gonadal somatic cell-derived factor (Gsdf), 9 genes had relatively high expression in ovary of XX-F, including bone morphogenetic protein 15 (Bmp15), cytochrome P450 family 19 subfamily A member 1a (Cyp19a1a) and forkhead transcription factor gene 2 (Foxl2). The results of qRT-PCR validation were consistent with the results of the RNA-seq, illustrating that Amhr2, Amh, Dmrt1 and Gsdf genes were related to the regulation of testicular development, and Bmp15, Cyp19a1a and Foxl2 genes were related to ovarian development regulation in C. argus. KEGG pathway enrichment analysis showed that most DEGs were enriched in the pathways related to sex differentiation and metabolism, such as mitogen-activated protein kinase (MAPK), wingless-type MMTV integration site family (Wnt) and transforming growth factor-β (TGF-β) signaling pathway. In this study, a large number of candidate genes and signaling pathways related to sex differentiation and gonadal development were screened out, which would provide preliminary basis and data support for further studies on the molecular mechanism of sex differentiation and gonadal development and the gene function of the sex-determining genes in C. argus.
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