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Isolation and Characterization of Spermatogenic Cells from Cattleyak,Cattle & Yak,and Cellular microRNA Expression Profiling

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Isolation and Characterization of Spermatogenic Cells from Cattleyak,Cattle & Yak,and Cellular microRNA Expression Profiling
论文目录
 
ABSTRACT第1-7页
摘要第7-13页
1 Introduction第13-16页
2 Literature review第16-36页
  2.1 Yak,cattle, cattleyak and significance of the hybridization第16-19页
    2.1.1 Yak第16-17页
    2.1.2 Cattle第17-18页
    2.1.3 Cattleyak第18-19页
  2.2 Approaches for spermatogenic cells isolation第19-21页
    2.2.1 Cell isolation by using Surface antigen/ immunophenotype第20页
    2.2.2 Cell isolation by using biological properties第20页
    2.2.3 Cell isolation by using physical properties第20-21页
  2.3 Brief introduction of microRNAs (miRNAs)第21-23页
    2.3.1 Biogenesis and mechanism of action of miRNAs第22-23页
  2.4 Spermatogenesis and microRNAs involvement第23-32页
    2.4.1 MicroRNAs and primordial germ cells第25-26页
    2.4.2 MicroRNAs and spermatogonial stem cells第26-28页
    2.4.3 MicroRNAs and differentiation of spermatogonia第28-29页
    2.4.4 MicroRNAs and meiosis of spermatocytes第29-31页
    2.4.5 MicroRNAs and spermiogenesis第31-32页
  2.5 Research background on cattleyak infertility第32-34页
    2.5.1 Research objectives第34页
  2.6 Experimental layout第34-36页
3 Materials and methods第36-61页
  3.1 Ethical statement for samples collection第36页
  3.2 TUNEL-POD analysis第36-39页
    3.2.1 Samples第36-37页
    3.2.2 Experimental regents and equipments第37-39页
    3.2.3 Protocol第39页
  3.3 Isolation and characterization of spermatogenic cells第39-51页
    3.3.1 Samples第39页
    3.3.2 Required equipments and chemicals第39-42页
    3.3.3 Required solutions第42页
    3.3.4 Required primers for (PCR) Polymerize Chain Reaction第42-43页
    3.3.5 Methodology for cell isolation第43-49页
    3.3.6 Methodology for identification of isolated cells第49-51页
  3.4 Cellular microRNAs expression profiling第51-55页
    3.4.1 Samples第51页
    3.4.2 Experimental reagents and instruments第51-52页
    3.4.3 MiRNA microarrays protocol第52-55页
  3.5 Bioinformatics analysis第55-58页
    3.5.1 In silico target genes prediction for DE miRNAs第56-57页
    3.5.2 GO and KEGG enrichment of the targeted genes第57-58页
  3.6 RT-Q PCR validation第58-60页
    3.6.1 Samples and required equipments第58页
    3.6.2 Required primers第58-59页
    3.6.3 Protocol of RT-Q PCR第59-60页
  3.7 Statistical analysis第60-61页
4 Results and discussion第61-99页
  4.1 Results第61-88页
    4.1.1 Cell apoptosis in cattleyak compared with yak第61-64页
    4.1.2 Morphological characterization of spermatogenic cells第64-67页
    4.1.3 Phenotypic characterization of spermatogenic cells第67-70页
    4.1.4 MicroRNAs expression profile in spermatogenic cells第70-77页
    4.1.5 Targeted genes for differentially expresses miRNAs第77-81页
    4.1.6 GO and KEGG enrichment of the targeted genes第81-87页
    4.1.7 RT q-PCR validation results第87-88页
  4.2 Discussion第88-99页
    4.2.1 Isolation and characterization of spermatogenic cells第88-93页
    4.2.2 Cellular microRNA expression profiles and cell apoptosisanalysis第93-99页
5 Conclusion第99-101页
  5.1 Suggestions第100-101页
6 Acknowledgement第101-103页
Special thank第102-103页
7 References第103-119页
Dedicated to my parents第119-120页
8 Appendix第120-139页
  8.1 Table. S1 Top 3/239 listed GO term BP, MF, CC of targetedgenes for DE miRNAs of CY vs CL第120-127页
  8.2 Table. S2 Top 3/243 listed GO term BP, MF, CC of targetedgenes for DE miRNAs of CY vs YK第127-135页
  8.3 Table. S3 KEGG top 5/35 listed pathways of targeted genes forDE miRNAs of CY vs CL第135-137页
  8.4 Table. S4 KEGG top 5/46 listed pathways of targeted genes forDE miRNAs of CY vs YK第137-139页
9 Publications第139页

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犏牛 雄性不育 生精细胞 生精细胞分离 STA-PUT法 MicroRNA测序 生物信息学分析
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