Investigation of Cardiac Gene and Protein Expressions Associ

发布时间：2021-03-05 05:16

　　青藏高原是全世界海拔最高的地区,有“世界屋脊”之称,平均海拔约为4000-5000米。高海拔物种由于在缺氧、高辐射、寒冷和生物产能缺乏的恶劣环境中生存并明显受到这些外界因素的影响,进而使其成为比较进化分析的理想研究对象。缺氧适应性是一个复杂的属性,多种因素参与其中。高海拔地区的牦牛很好地适应了低氧环境,但是到目前为止,与其相关的心肌基因和蛋白表达调控机制的研究还未见报道。青藏高原地区的动物具有较强的心脏功能,与缺氧适应性相关的基因随着机体体循环增强而表达,并参与保护心脏细胞和组织结构。转录组和蛋白质组分析都曾用于筛选牦牛和其他家养物种肌肉生长脂质沉积的功能基因和蛋白质。为了揭示牦牛缺氧适应的分子机制,本研究采用RNA测序和iTRAQ技术对牦牛和黄牛进行心肌比较转录组学和蛋白组学分析。为了系统理解牦牛心肌组织中基因表达变异与其缺氧适应性的关系,RNA测序分析后从牦牛和黄牛心肌组织中鉴定得到20646个差异表达基因（DEGs）。其中,GSTA2、AHCYL2、SOD1、Idh1、GATM、HK3和BCAA基因与高海拔适应性相关,参与细胞内钙释放、缺氧下HIF-1α的稳定、通过改变异柠檬酸盐... 

【文章来源】：西南科技大学四川省

【文章页数】：129 页

【学位级别】：博士

【文章目录】：
摘要
ABSTRACT
1 Chapter Introduction and research backgrounds
    1.1 Introduction
    1.2 Research backgrounds
    1.3 Objectives of this study
2 Chapter Review of literature
    2.1 Review of the literature
        2.1.1 Heart/ Cardia
        2.1.2 Cardia genesis
        2.1.3 The origin of cardio myocytes, outflow tract and smooth muscle cells
        2.1.4 Septum Formation
        2.1.5 Cardiac looping
        2.1.6 Convergence
        2.1.7 Wedging
        2.1.8 The venous pole of heart
        2.1.9 Atrial septation
        2.1.10 Formation of Ventricular
    2.2 Cardiac conduction system （CCS）
    2.3 Transcriptional taxonomy in Cardiac Conduction System Development
        2.3.1 HCN4
        2.3.2 Nkx2-5
        2.3.3 Tbx3
        2.3.4 Pitx2
    2.4 Cardiac Energy Metabolism
    2.5 Fatty Acid Metabolism
    2.6 Glucose Metabolism
    2.7 Interaction amongst Fatty acids and Glucose Metabolism
    2.8 Cardiac Energy Metabolism and Efficiency
    2.9 Conclusions
3 Chapter Comparative analysis of myocardium transcriptome identified genes associated with high-altitude adaptation in Yak
    3.1 Introduction
    3.2 Materials and methods
        3.2.1 Animals and myocardium sample collection
        3.2.2 Morphological analysis of heart in Yak and Cattle
        3.2.3 Histological analysis of myocardium samples from yak and cattle
        3.2.4 RNA extraction and cDNA library preparation
        3.2.5 Data Processing, and assembly of yak and cattle transcriptome
        3.2.6 Screening of DEGs and Gene expression analysis
        3.2.7 GO enrichment analysis of DEGs
        3.2.8 KEGG Pathway enrichment analysis of DEGs
        3.2.9 RT-PCR and qRT-PCR Validation of mRNA-Sequence data
        3.2.10 Statistical analysis
    3.3 Results
        3.3.1 Morphological analysis of heart in Yak and Cattle
        3.3.2 Histological characteristics of myocardium in Yak vs cattle
        3.3.3 DNA sequencing, tag mapping, and patterns of gene expression
        3.3.4 DEGs between yak and cattle myocardial transcriptome
        3.3.5 GO enrichment analysis of DEGs
        3.3.6 KEGG Pathway enrichment analysis of myocardial DEGs
    3.4 Discussion
        3.4.1 Morphological & Histological characteristic analysis of heart in Yak vs
        3.4.2 KEGG functional enrichment analysis of DEGs in Yak
        3.4.3 KEGG functional enrichment analysis of DEGs in Yak
    3.5 Conclusion
4 Chapter Comparative iTRAQ proteomics identified myocardium proteins associated with hypoxia ofyak
    4.1 Introduction
    4.2 Material and Methods
        4.2.1 Sample collection
        4.2.2 Protein Preparation
        4.2.3 iTRAQ Labeling and SCX fractionation
        4.2.4 LC-ESI-MS/MS analysis
        4.2.5 Bioinformatics
        4.2.6 COG and GO analysis for all identified proteins
        4.2.7 GO and KEGG pathway enrichment analysis of DEPs and string analysis
    4.3 RESULTS
        4.3.1 Protein Identification
        4.3.2 DEPs between yak and cattle myocardial proteome
        4.3.3 GO analysis of DEPs
        4.3.4 KEGG pathway enrichment and Protein-protein interaction by STRINGnetwork68
    4.4 Discussion
        4.4.1 DEPs in yak associated with myocardial development
        4.4.2 DEPs in yak associated with mitochondrial functions
        4.4.3 DEPs in yak in response to the immune system
    4.5 Conclusion
5 Chapter Summary Conclusion and Recommendations for Future Research
    5.1 Summary
    5.2 General Conclusion and Recommendations for Future Research
6 Acknowledgments Special thanks
Special thanks
7 References
8 Appendix A
    8.1 S1 Primers information for RT-PCR and Q-PCR validation
    8.2 S2 List of top enriched GO terms for biological function
    8.3 S3 List of KEGG enriched pathway of DEGs in YK and CL
    8.4 S4 List of top three enriched GO terms for biological process, Cellular componentand molecular functions in myocardium
    8.5 S5 List of top ten KEGG enriched pathway of DEPs in Yak and Cattle
    8.6 S6 The top 10 significantly enriched KEGG pathways according to protein-protein interaction in STRING network analysis
9 Publications or research achievement



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