樱桃谷鸭胚胎期毛囊发育miRNA表达谱及相关差异表达基因的研究

发布时间：2018-03-01 17:37

本文关键词： 樱桃谷鸭 microRNA 毛囊高通量测序　出处：《安徽农业大学》2016年硕士论文　论文类型：学位论文

【摘要】：羽绒性状是鸭重要的经济性状之一。羽毛发育受多种因素的影响,其中遗传因素发挥重要作用。miRNA可通过调节基因表达进而影响羽毛发育。本研究通过对樱桃谷鸭胚胎期不同发育阶段毛囊发育miRNA表达谱的研究,从mi RNA水平上揭示影响羽毛发育遗传调控的关键分子,为鸭羽毛发育的遗传机理提供科学理论基础。试验选取樱桃谷鸭胚胎期E11、E15、E20和出雏1d皮肤样品(每时期3个重复),建立各时期mi RNA表达谱文库,同时选取差异表达miRNA进行qRT-PCR验证,miRNA与羽毛发育有关基因的相关性采用SAS9.4简单相关性分析。与鸡miRBase数据库比对,共获得已知miRNAs 337个,其中差异表达的miR NA为216个。新miRNAs 93个。根据表达特征,可将差异表达miRNA分为8种类型。根据miRNA在测序数据库中的表达量,筛选出7个高表达miRNA,可能与羽毛发育调控相关,KEGG Pathway分析共获得12条信号通路,其中包括Notch信号通路、磷脂酰肌醇信号通路等一些参与毛囊发生发育的信号通路。由此可见,Notch等一些明确参与毛囊发育的信号通路和甘油酯代谢等一些脂质合成代谢途径可能与羽毛的品质相关。为验证数据库的准确性,选定已知数据库中两个差异表达的miRNAs和一个差异表达的新miRNA通过qRT-PCR表达分析不同时期的相对表达水平,定量结果与mi RNA表达谱结果具有较高的一致性。为探索数据表达结果,我们将已选中的三个mi RNAs与相关基因(Fzd3,Wnt8a,Lrp6,and Birc6)进行Pearson相关系数分析(r-v alues)。miR-30a-5p和miR-203a表达量与Fzd3表达量呈显著负相关(r=-0.663,P=0.0004;r=-0.640,P=0.0008)。此外,gga-miR-new-20表达量与Wnt8a和Birc6表达量呈显著负相关(r=-0.421,P=0.040;r=-0.778,P0.0001)。由此可见,Wnt信号途径在胚胎发育早期即开始参与毛囊生长调控过程。
[Abstract]:Feather traits are one of the important economic traits in duck. Feather development is affected by many factors. Genetic factors play an important role. MiRNAs can affect feather development by regulating gene expression. In this study, miRNA expression profiles of hair follicles in different stages of embryonic development of Cherry Valley ducks were studied. The key molecules affecting the genetic regulation of feather development were revealed at the mi RNA level. To provide a scientific theoretical basis for the genetic mechanism of duck feather development, we selected E11E15 E20 and 1 day skin samples of Cherry Valley Duck at embryonic stage (3 repeats per stage) to establish the expression library of mi RNA in each stage. At the same time, differential expression miRNA was selected to verify the correlation between miRNAs and genes related to feather development using SAS9.4 simple correlation analysis. Compared with chicken miRBase database, a total of 337 known miRNAs were obtained. Among them, 216 miRNA were differentially expressed and 93 new miRNAs. According to the expression characteristics, the differentially expressed miRNA could be divided into 8 types. According to the expression amount of miRNA in sequencing database, Seven highly expressed miRNAs were screened, and 12 signal pathways were obtained by KEGG Pathway analysis, including Notch signaling pathway, which may be related to the regulation of feather development. Some signal pathways such as phosphatidylinositol signaling pathway are involved in hair follicle development. It can be seen that some signal pathways such as Notch and glycerol metabolism may be related to feather. Quality correlation. To verify the accuracy of the database, Two differentially expressed miRNAs and a new differentially expressed miRNA in known databases were selected to analyze the relative expression levels at different periods by qRT-PCR expression. The quantitative results were in good agreement with the results of miRNA expression profile. Pearson correlation coefficient analysis was performed between the three selected mi RNAs and the related gene Fzd3 / Wnt8aFrp6 and Birc6). There was a significant negative correlation between the expression of r-v alues).miR-30a-5p and miR-203a and the expression of Fzd3. It can be seen that the expression of r-v alues).miR-30a-5p and miR-203a is negatively correlated with the expression of Fzd3, and the expression of new-gga-miR-20 is negatively correlated with the expression of Wnt8a and Birc6, and it can be seen that there is a significant negative correlation between the expression of r-v alues).miR-30a-5p and miR-203a and the expression of Fzd3. It can be seen that there is a significant negative correlation between the expression of new-gga-miR-20 and the expression of Wnt8a and Birc6. The Wnt signaling pathway takes part in the regulation of hair follicle growth at the early stage of embryonic development.
【学位授予单位】：安徽农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S834;Q78

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