In previous study, we reported the identification, tissue distribution, and the roles of Spdsx played in the testis, androgenic gland, and ovary in Scylla paramamosain. Here, we primally identify its potential target genes in the ovary with RNAi and RNA-Seq technology. By comparing the transcriptome data of two groups (ovaries that injected with dsRNA for EGFP and Dsx), we found that 6520 Unigenes were differentially expressed, including a plenty of conserved crucial genes involved in ovarian development, such as vitellogenin (vtg), vtg receptor (vtgR), apolipoprotein D, adenylate cyclase 3, adenylate cyclase 5, cyclin A, cyclin B, and cell division cycle 2 (cdc2). In addition, these DEGs were also enriched in pathways related to ovary development, including PI3K-Akt signaling pathway, MAPK signaling pathway, insulin signaling pathway, Wnt signaling pathway, relaxin signaling pathway, estrogen signaling pathway, progesterone-mediated oocyte maturation, ovarian steroidogenesis, and oocyte meiosis. Moreover, several genes were selected for qRT-PCR to validate the accuracy of the bioinformatic result. According to current transcriptome result, we speculate that the Spdsx is a crucial regulator of ovary development in S. paramamosain. To the best of our knowledge, the current study was the first report about dsx function through comparative transcriptome analysis in crustacean species, which not only identified relevant genes and pathways involved in ovarian development of S. paramamosain, but also shed light on the regulatory mechanisms of dsx at the molecular level in crustacean.
Abayed FA, Manor R, Aflalo ED, Sagi A (2019) Screening for Dmrt genes from embryo to mature Macrobrachium rosenbergii prawns. Gen Comp Endocrinol 282:113205
Burtis KC, Baker BS (1989) Drosophila doublesex gene controls somatic sexual differentiation by producing alternatively spliced mRNAs encoding related sex-specific polypeptides. Cell 56:997–1010
[DOI:
10.1016/0092-8674(89)90633-8]
Carnevali O, Cionna C, Tosti L, Lubzens E, Maradonna F (2006) Role of cathepsins in ovarian follicle growth and maturation. Gen Comp Endocrinol 146:195–203
[DOI:
10.1016/j.ygcen.2005.12.007]
Chen T, Ren C, Jiang X, Zhang L, Li H, Huang W, Hu C (2018) Mechanisms for type-II vitellogenesis-inhibiting hormone suppression of vitellogenin transcription in shrimp hepatopancreas: crosstalk of GC/cGMP pathway with different MAPK-dependent cascades. PLoS One 13:e0194459
Du J, Liu Y, Song C, Cui Z (2019) Discovery of sex-related genes from embryonic development stage based on transcriptome analysis in Eriocheir sinensis. Gene 710:1–8
Duan J, Xu H, Ma S, Guo H, Wang F, Zhang L, Zha X, Zhao P, Xia Q (2014) Ectopic expression of the male BmDSX affects formation of the chitin plate in female Bombyx mori. Mol Reprod Dev 81:240–247
Esteban-Lopez M, Agoulnik AI (2020) Diverse functions of insulin-like 3 peptide. J Endocrinol 247:R1–R12
Farhadi A, Fang S, Zhang Y, Cui W, Fang H, Ikhwanuddin M, Ma H (2021) The significant sex-biased expression pattern of Sp-Wnt4 provides novel insights into the ovarian development of mud crab (Scylla paramamosain). Int J Biol Macromol 183:490–501
[DOI:
10.1016/j.ijbiomac.2021.04.186]
Gempe T, Beye M (2011) Function and evolution of sex determination mechanisms, genes and pathways in insects. BioEssays 33:52–60
[DOI:
10.1002/bies.201000043]
Guo L, Xie W, Liu Y, Yang Z, Yang X, Xia J, Wang S, Wu Q, Zhang Y (2018) Identification and characterization of doublesex in Bemisia tabaci. Insect Mol Biol 27:620–632
Haraguchi S, Ikeda N, Abe M, Tsutsui K, Mita M (2016) Nucleotide sequence and expression of relaxin-like gonad-stimulating peptide gene in starfish Asterina pectinifera. Gen Comp Endocrinol 227:115–119
[DOI:
10.1016/j.ygcen.2015.06.017]
Huang X, Feng B, Huang H, Ye H (2017) In vitro stimulation of vitellogenin expression by insulin in the mud crab, Scylla paramamosain, mediated through PI3K/Akt/TOR pathway. Gen Comp Endocrinol 250:175–180
Jordan BK, Shen JH, Olaso R, Ingraham HA, Vilain E (2003) Wnt4 overexpression disrupts normal testicular vasculature and inhibits testosterone synthesis by repressing steroidogenic factor 1/beta-catenin synergy. Proc Natl Acad Sci USA 100:10866–10871
Li S, Li F, Yu K, Xiang J (2018) Identification and characterization of a doublesex gene which regulates the expression of insulin-like androgenic gland hormone in Fenneropenaeus chinensis. Gene 649:1–7
[DOI:
10.1016/j.gene.2018.01.043]
Lv J, Li R, Su Z, Gao B, Ti X, Yan D, Liu G, Liu P, Wang C, Li J (2022) A chromosome-level genome of Portunus trituberculatus provides insights into its evolution, salinity adaptation and sex determination. Mol Ecol Resour 22:1606–1625
[DOI:
10.1111/1755-0998.13564]
Mani T, Subramaniya BR, Chidambaram Iyer S, Sivasithamparam ND, Devaraj H (2019) Modulation of complex coordinated molecular signaling by 5HT and a cocktail of inhibitors leads to ovarian maturation of Penaeus monodon in captivity. Mol Reprod Dev 86:576–591
[DOI:
10.1002/mrd.23135]
Mine S, Sumitani M, Aoki F, Hatakeyama M, Suzuki MG (2021) Effects of functional depletion of doublesex on male development in the sawfly Athalia Rosae. Insects 12. https://doi.org/10.3390/insects12100849
Mita M, Elphick MR, Katayama H (2020a) Effect of chimeric relaxin-like gonad-stimulating peptides on oocyte maturation and ovulation in the starfish Asterias rubens and Aphelasterias japonica. Gen Comp Endocrinol 287:113351
[DOI:
10.1016/j.ygcen.2019.113351]
Ministry of Agriculture and Fisheries, The China Society of Fisheries. (2020). China Fishery Statistical Yearbook 2021. China Agriculture Press, Beijijng. (in Chinese)
Mita M, Matsubara S, Osugi T, Shiraishi A, Wada A, Satake H (2020b) A novel G protein-coupled receptor for starfish gonadotropic hormone, relaxin-like gonad-stimulating peptide. PLoS One 15
Mita M, Osugi T, Matsubara S, Kawada T, Satake H, Katayama H (2021) A relaxin-like gonad-stimulating peptide identified from the starfish Astropecten scoparius. Mol Reprod Dev 88:34–42
[DOI:
10.1002/mrd.23444]
Mita M, Yoshikuni M, Ohno K, Shibata Y, Paul-Prasanth B, Pitchayawasin S, Isobe M, Nagahama Y (2009) A relaxin-like peptide purified from radial nerves induces oocyte maturation and ovulation in the starfish, Asterina pectinifera. Proc Natl Acad Sci USA 106:9507–9512
Nakata M, Kikuchi Y, Iwami M, Takayanagi-Kiya S, Kiya T (2021) Identification and characterization of sexually dimorphic neurons that express the sex-determining gene doublesex in the brain of silkmoth Bombyx mori. Insect Biochem Mol Biol 129:103518
Shukla JN, Nagaraju J (2010) Doublesex: a conserved downstream gene controlled by diverse upstream regulators. J Genet 89:341–356
Steegborn C (2014) Structure, mechanism, and regulation of soluble adenylyl cyclases - similarities and differences to transmembrane adenylyl cyclases. Biochem Biophys Acta 1842:2535–2547
Vainio S, Heikkila M, Kispert A, Chin N, Mcmahon AP (1999) Female development in mammals is regulated by Wnt-4 signalling. Nature 397:405–409
[DOI:
10.1038/17068]
Wan H, Zhong J, Zhang Z, Sheng Y, Wang Y (2022) Identification and functional analysis of the doublesex gene in the mud crab Scylla paramamosain. Comp Biochem Physiol 266:111150
Wan H, Zhong J, Zhang Z, Zou P, Zeng X, Wang Y (2021) Discovery of the Dmrt gene family members based on transcriptome analysis in mud crab Scylla paramamosain. Gene 784:145576
[DOI:
10.1016/j.gene.2021.145576]
Yuzawa T, Matsuoka M, Sumitani M, Aoki F, Sezutsu H, Suzuki MG (2020) Transgenic and knockout analyses of Masculinizer and doublesex illuminated the unique functions of doublesex in germ cell sexual development of the silkworm. Bombyx Mori BMC Development Biology 20:19
[DOI:
10.1186/s12861-020-00224-2]
Zheng J, Cai L, Jia Y, Chi M, Cheng S, Liu S, Li F, Gu Z (2020) Identification and functional analysis of the doublesex gene in the redclaw crayfish. Cherax Quadricarinatus Gene Expr Patterns 37
Zhong Y, Zhao W, Tang Z, Huang L, Zhu X, Liang X, Yan A, Lu Z, Yu Y, Tang D, Wang D, Lu Z (2021) Comparative transcriptomic analysis of the different developmental stages of ovary in red swamp crayfish Procambarus clarkii. BMC Genomics 22:199
[DOI:
10.1186/s12864-021-07537-x]
