摘要
为探究XX雌性和XY雄性金钱鱼(Scatophagus argus)下丘脑中的差异表达基因及雌激素(17β-Estradiol,E2)在体注射对XY雄鱼下丘脑中基因表达的影响,开展了转录组测序分析,包括测序数据质控、基因功能注释,差异基因筛选、鉴定和差异基因功能富集分析等,并利用实时定量PCR(Real-time quantitative PCR,qPCR)检测了金钱鱼下丘脑中相关基因的表达。金钱鱼下丘脑转录组共测得Clean reads 275833710,测序质量30(Q30)大于95%,GC含量值大于48%。其中,Ctrl-XX-H vs.Ctrl-XY-H组,共筛选和鉴定了91个差异表达基因,包括36个上调基因和55个下调基因。在Ctrl-XY-H vs.E2-XY-H组,筛选和鉴定了28个差异表达基因,包括11个上调基因和17个下调基因。GO和KEGG富集分析发现,差异表达基因主要显著富集在细胞、单生物过程,膜、膜组分,结合等生物学功能及泛醌和其他萜类-醌生物合成及类固醇激素生物合成通路等。转录组和qPCR结果表明,XX和XY金钱鱼下丘脑中prl、ccna1和hsp90aa1.1等生殖相关基因的表达具有明显的性别差异,但sbGnRH、sGnRH和cGnRH等生殖轴相关基因,ghrh、ghrhr和sst1、3、5和6等生长轴相关基因的表达没有明显差异。E2在体注射后,雄鱼下丘脑中cyp19a1b和greb1等基因的表达显著上调,而prl的表达显著下调。以上结果表明,雌激素可通过反馈作用调节下丘脑中部分基因的表达,参与调控金钱鱼性腺发育和配子发生过程。
Hypothalamus are the important endocrine organs,which play key roles in regulating body growth,reproduction and hormone activity in other endocrine glands and organs.Estrogen,an important sex steroid hormone,is mainly synthesized and secreted in the gonads.It could regulate the reproduction and growth related genes expression in hypothalamus through feedback,and then participate in the regulation of corresponding physiological activities.Spotted scat(Scatophagus argus)is a good model to investigate the sexual growth dimorphism with a faster growth in females than males.The effect of 17β-estradiol(E2)injection in vivo on genes expression in hypothalamus of XY male spotted scat and the differences in genes expression between XY males and XX females remain unclear.In order to explore gene expression difference between XY male and XX female spotted scat hypothalamus and effect of in vivo injection in XY males,transcriptome analysis of hypothalamus were carried out,including sequencing data quality control,gene function annotation,differentially expressed gene(DEGs)screening and identification and pathway enrichment analysis,then real-time quantitative PCR(qPCR)was performed to detect genes expression.A total of 275833710 Clean reads were sequenced in this study,and the Q30 and GC value were more than 95%and 48%,respectively.In Ctrl-XX-H vs.Ctrl-XY-H group,91 DEGs were screened and identified successfully,including 36 up-regulated and 55 down-regulated genes.In Ctrl-XX-H vs.E2-XY-H group,28 DEGs were screened and identified,including 11 up-regulated and 17 down-regulated genes.GO and KEGG pathway analysis showed that DEGs was significantly enriched in biological processes such as cells,single cell,membrane,membrane components,and signal pathways such as ubiquinone and other terpene quinone biosynthesis,steroid hormone biosynthesis.By transcriptome and qPCR analyses,the expression of nrip1b,prl,hspa8b,snx21,cep63,ccna1,hsp90aa1.1,slc1a5,zar1,slc25a1b and loc124068175,were significantly different between XX and XY spotted scat,while the reproductive axis genes(sbGnRH,sGnRH and cGnRH)and growth axis genes(ghrh,sst1,sst3,sst5 and sst6)expression showed no significant differences.In XY male fish,six hours after E2 injection,the expression of cdc42ep1a,cyp19a1b,greb1,kank1a,snx21,and tmem30c in hypothalamus were significantly up-regulated,while prl expression was significantly down-regulated.Above results suggest that E2 might regulate some reproduction related genes expression in hypothalamus through feedback effects,which plays the critical roles in gametogenesis and reproduction regulation.
作者
潘书慧
周彤
袁嘉慧
罗琼
秦若彤
茹笑影
黄洋
石红娟
李广丽
PAN Shu-Hui;ZHOU Tong;YUAN Jia-Hui;LUO Qiong;QIN Ruo-Tong;RU Xiao-Ying;HUANG Yang;SHI Hong-Juan;LI Guang-Li(Fisheries College of Guangdong Ocean University,Key Laboratory of Aquaculture in South China Sea for Aquatic Economic Animal of Guangdong Higher Education Institutes,Zhanjiang 524088,China;Guangdong Province Famous Fish Reproduction Regulation and Breeding Engineering Technology Research Center of Engineering Technology Research Center,Zhanjiang 524088,China)
出处
《水生生物学报》
CAS
CSCD
北大核心
2023年第8期1195-1210,共16页
Acta Hydrobiologica Sinica
基金
国家自然科学基金(32002367)
国家重点研发计划“蓝色粮仓科技创新”专项(2018YFD0901203)
广东海洋大学博士启动项目(R20052)资助。
