Full-Length Transcriptomic Analysis of Chicken Pituitary Reveals Candidate Genes for Testicular Trait
马荆鄂 1熊信威 1周敏 1吴斯琪 1韩甜 1饶友生 1王樟凤 1许继国1
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作者信息
1. 南昌师范学院生物技术研究院/家禽遗传改良江西省重点实验室,南昌 330032
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摘要
[背景]睾丸是雄性动物重要生殖器官,具有产生精子和分泌雄激素的功能,睾丸质量与繁殖性能有直接联系.睾丸性状受下丘脑-垂体-性腺轴多基因共同调控,其分子机理尚不清楚.宁都黄鸡睾丸性状变异系数大,繁殖性能一致性差.[目的]通过对不同睾丸性状个体垂体组织进行全长转录组测序,挖掘候选基因以及关键信号通路,探究影响睾丸性状的遗传基础,以期为发展地方鸡种睾丸性状的选种选育提供依据.[方法]以 70 只 22 周龄宁都黄公鸡为研究对象,分为大睾丸组和小睾丸组,从两组中分别选取 3 个个体,构建垂体组织的全长转录本表达谱,筛选高表达量及两组个体差异表达基因或转录本,对差异基因或转录本进行功能富集分析.随机选取 6 个基因,进行基因表达水平验证.参考已知垂体细胞的标记基因,筛选与 8 种细胞标记基因对应的差异表达基因或转录本,并分析这些差异基因或转录本以及高表达量基因或转录本与睾丸性状相关性.[结果]在 6 个宁都黄鸡垂体组织中,检测到 21 834 个基因,29 355 个全长转录本.其中,332 个基因及 229 个转录本在两组个体中差异表达.KEGG 通路分析结果显示,这些差异基因或转录本主要富集于Apelin信号、甘油磷脂代谢等通路.一共发现 5 个促性腺细胞标记基因或其对应转录本与部分睾丸性状显著相关,包括NFASC(neurofascin)及其对应转录本 XM_015298904.2、TESC(tescalcin)及其对应转录本XM_025155510.1、FSHB(follicle stimulating hormone beta)转录本 ONT.20974.4、RLN3(relaxin 3)、NDUFB2(NADH:ubiquinone oxidoreductase subunit B2)转录本ONT.1176.4.1 个催乳素细胞标记基因EGR1(early growth response 1),2 个滤泡星状细胞标记基因TMSB4X(thymosin beta 4,X-linked)、C1H12ORF57(chromosome 1 open reading frame),1 个促甲状腺细胞标记基因CHGB(chromogranin B)均与部分睾丸性状显著相关.2 个高表达量基因TMSB4X、HSP90AA1(heat shock protein 90 alpha family class A member 1)与部分睾丸性状显著相关.FSHB、EGR1均为基因互作网络中节点基因.[结论]经初步功能分析,EGR1、TMSB4X、FSHB、RLN3为睾丸性状调控的关键基因.MAPK 信号通路、神经活性配体-受体相互作用、GnRH信号通路为睾丸性状调控的关键基因通路.
Abstract
[Background]Testis is an important reproductive organ of male animals,which has the function of producing sperm and secreting androgen.The quality of testis is directly related to reproductive performance.Testicular traits are regulated by multiple genes in the hypothalamic-pituitary-gonadal axis,but the molecular mechanism is still unclear.The variation coefficient of testicular traits was large in Ningdu yellow chicken,with poor reproductive performance consistency.[Objective]In order to explore candidate genes and key signaling pathways,full-length transcriptome sequencing was performed on pituitary tissues of six individuals with different testicular traits.The selection and breeding of testis traits was expected to develop local chicken breeding with these founding genetic basis.[Method]A total of seventy 22-week-old Ningdu yellow roosters were studied.Six individuals were randomly selected,including three individuals with large testicular and others with small testicular.Full-length transcript expression profile was constructed on pituitary tissue for each individual.Then the genes or transcripts were selected with high expression or different expression between two groups of individuals.Gene function enrichment analysis was also performed for these differentially expressed genes or transcripts.Six genes were randomly selected to verify the gene expression level.Referred to known marker genes in pituitary cells,screening the differentially expressed genes or transcripts were carried out corresponding to the marker genes in the eight kinds of cells.The correlation was analyzed between testicular traits and these genes or transcripts and also those with high expressional level.[Result]In six pituitary tissues,21 834 genes and 29 355 full-length transcripts were detected.Among them,332 genes and 229 transcripts were differentially expressed between the two groups.KEGG pathway analysis showed that differentially expressed genes or transcripts were mainly concentrated in apelin signaling pathway,and glycerophospholipid metabolism.Five marker genes of the gonadotropin cell were identified related to part of testicular traits,including NFASC(neurofascin)and its transcript XM_015298904.2,TESC(tescalcin)and its transcript XM_025155510.1,ONT.20974.4(one transcript of follicle stimulating hormone beta),RLN3(relaxin 3),ONT.1176.4(one transcript of NADH:ubiquinone oxidoreductase subunit B2).Four genes were also identified related to part of testicular traits,including EGR1(early growth response 1)as one of the marker genes in the lactotrophs,TMSB4X(thymosin beta 4,X-linked)and C1H12ORF57(chromosome 1 open reading frame)as the marker genes of the folliculo-stellate cells,CHGB(chromogranin B)as one of the marker genes in the thyrotrophs.Two highly expressed genes were identified related to part of testicular traits,including TMSB4X and HSP90AA1(heat shock protein 90 alpha family class A member 1).FSHB and EGR1 were found as node genes in the gene interaction network.[Conclusion]Preliminary functional analysis showed that EGR1,TMSB4X,FSHB and RLN3 were the key genes in testicular trait regulation.MAPK,neuroactive ligand-receptor interaction and GnRH signaling pathway played the key role in the regulation of testicular traits.
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