绍兴鸭血清生殖激素及下丘脑开产相关基因的鉴定分析

Identification and analysis of serum reproductive hormones and genes related to laying in hypothalamus of Shaoxing Ducks

    摘要
  • 摘要: 【目的】比较开产和未开产绍兴鸭血清中的生殖激素水平，挖掘下丘脑组织中与开产性状相关的候选基因，为利用分子标记进行鸭开产性状选育打下理论基础。【方法】以105和128日龄开产与未开产绍兴鸭为研究对象，采用放射免疫测定法（RIA）测定血清中雌二醇（E2）、卵泡刺激素（FSH）和黄体生成素（LH）的浓度，利用转录组测序技术筛选105日龄开产与未开产绍兴鸭下丘脑组织中的差异表达基因（DEGs），通过GO和KEGG数据库对DEGs进行GO功能注释和KEGG信号通路富集分析，并以实时荧光定量PCR验证转录组数据的准确性。【结果】在105和128日龄，开产绍兴鸭血清的E2浓度极显著高于未开产绍兴鸭（P<0.01，下同），LH浓度显著低于未开产绍兴鸭（P<0.05，下同）；在128日龄，开产绍兴鸭血清的FSH浓度极显著低于未开产绍兴鸭。基于转录组测序，从105日龄开产与未开产绍兴鸭的下丘脑组织共筛选出617个DEGs，其中上调表达的DEGs有91个、下调表达的DEGs有526个。GO功能注释分析显示，DEGs主要注释在神经系统发育、激素活性和细胞迁移等功能条目上； KEGG信号通路富集分析发现，神经活性配体—受体相互作用和内吞作用可能是与早熟性能相关的信号通路。实时荧光定量PCR检测结果显示，开产绍兴鸭下丘脑中的NPY、ITIH2和GDF10基因相对表达量较未开产绍兴鸭极显著上调，WIF1、HSP70和Ret基因相对表达量显著下调，而FASN基因的相对表达量无显著差异（P>0.05），与转录组测序结果一致。【结论】从绍兴鸭下丘脑组织筛选到的NPY、GnIH、TRH和HSP70等DEGs通过神经活性配体—受体相互作用和内吞作用等信号通路，作用于下丘脑—垂体—性腺轴，影响E2、FSH和LH等生殖激素的分泌与释放，进而调控绍兴鸭的开产时间。

     

    Abstract: 【Objective】The purpose of the study was to compare the levles of reproductive hormone in serum of laying and pre-laying Shaoxing Ducks and to screen the candidate genes related to laying traits in the hypothalamic tissue, so as to provide a theoretical basis for use of molecular markers for the selection of laying performance of ducks.【Method】 The 105-day-old and 128-day-old laying and pre-laying Shaoxing Ducks were selected as the research subjects. The concentrations of estrogen(E2),follicle stimulating hormone(FSH)and luteinizing hormone(LH)in serum were determined by radioimmunoassay(RIA). Differentially expressed genes(DEGs)of 105-day-old laying and pre-laying Shaoxing Ducks in the hypothalamus tissues were screened by transcriptome sequencing. GO functional annotation and KEGG signaling pathway enrichment analysis were performed on DEGs through GO and KEGG databases,and the accuracy of transcriptome data was verified by real-time fluorescence quantitative PCR.【Result】 At 105-day-old and 128-day-old,the concentrations of E2 in the serum of laying Shaoxing Ducks were extremely significantly higher than those of pre-laying Shaoxing Ducks(P<0.01,the same below),and the concentrations of LH in the serum of laying Shaoxing Ducks were significantly lower than those of pre-laying Shaoxing Ducks(P<0.05,the same below). At 128-day-old,the concentrations of FSH in the serum of laying Shaoxing Ducks were significantly lower than those of pre-laying Shaoxing Ducks. Based on transcriptome sequencing,a total of 617 DEGs were screened from the hypothalamic tissues of 105-day-old laying and pre-laying Shaoxing Ducks,among which 91 DEGs were up-regulated and 526 were down-regulated. GO functional annotation analysis showed that DEGs were mainly annotated on functional entries such as nervous system development, hormone activity and cell migration. KEGG signaling pathway enrichment analysis revealed that neuroactive ligandreceptor interaction and endocytosis may be signaling pathways related to precocious performance. The results of real-time fluorescence quantitative PCR showed that the relative expressions of NPY,ITIH2 and GDF10 genes in the hypothalamus of laying Shaoxing Ducks were extremely significantly up-regulated compared with those of pre-laying Shaoxing Ducks, and the relative espressions of WIF1,HSP70 and Ret genes in the laying Shaoxing Ducks were extremely significantly down-regulated than those of pre-laying Shaoxing Ducks,whereas there was no significant difference in the relative expression of FASN gene(P>0.05), which were basically consistent with the transcriptome sequencing results.【Conclusion】 The DEGs such as NPY,GnIH, TRH and HSP70 screened from the hypothalamus of Shaoxing Ducks act on the hypothalamus-pituitary-gonadal axis through signaling pathways such as neuroactive ligand-receptor interaction and endocytosis, and affect the secretion and release of reproductive hormones such as E2, FSH and LH, and then regulate the time of laying in Shaoxing Ducks.

     

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