17β-雌二醇注射雌性金钱鱼下丘脑转录组分析

Transcriptomic Analysis of Hypothalamus in Female Scatophagus argus after 17β-Estradiol Injection

【目的】通过转录组测序分析雌激素(17β-Estradiol,E_(2))对雌性金钱鱼下丘脑中基因表达的影响。【方法】E_(2)体注射后进行雌性金钱鱼下丘脑转录组测序,基因功能注释,差异基因筛选、鉴定,GO(Gene Ontology annotation)和KEGG(Kyoto Encyclopedia of Genes and Genomes)信号通路富集分析,并利用实时定量PCR(Real-time quantitative PCR,qPCR)检测E_(2)体注射后雌性金钱鱼下丘脑中相关基因的表达。【结果】雌性金钱鱼下丘脑转录组测序共测得Clean reads为190909756,注释到17687个基因,筛选和鉴定了22个差异表达基因,包括prl、erf3α、pgr和cyp19a1b等11个表达上调基因和chsy1、muc17l、prf1和hla-α等11个下调基因。通过KEGG功能富集分析发现差异基因涉及13个代谢通路。转录组和qPCR结果显示,与对照雌鱼相比,E_(2)注射组雌鱼下丘脑中prl、pgr、cyp19a1b和3β-HSDI的表达显著上调,而生长轴相关基因ghrh、sst1、sst3、sst5和sst6的表达未受影响。【结论】E_(2)通过反馈作用调节下丘脑中部分性类固醇激素合成通路相关基因的表达,但不影响生长相关基因的表达。

【Objectives】To study the mechanism of sexual dimorphism in growth and follicle development in female spotted scat after estrogen(17β-Estradiol,E_(2))injection.【Methods】The transcriptome sequencing of the hypothalamus was performed in female fish after E_(2) injection.Then,genes function annotation,differential genes screening,identification,Gene Ontology annotation(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment and differential genes expression by Real-time quantitative PCR(qPCR)was performed.【Results】A Total of 190909756 Clean reads and 17687 annotated genes were yield.Compared with the control hypothalamus,22 differential genes were screened and identified,of which 11 genes including prl,erf3α,pgr and cyp19a1b were up-regulated,11 genes including chsy1,muc17l,prf1 and hla-αwere down-regulated.KEGG functional enrichment analysis showed that the differential genes were found to be involved in 13 metabolic pathways.Consistently,qPCR and transcriptome analysis results revealed that prl,pgr,cyp19a1b and 3β-HSDI mRNA expressions were significantly up-regulated,while ghrh,sst1,sst3,sst5 and sst6 mRNA expressions showed no differences in hypothalamus of the E_(2) injection female fish,compared with the control fish.【Conclusion】E_(2) up-regulates the expression of some steroidogenesis-related genes in the hypothalamus through feedback,but does not affect growth-related genes.