| Abbreviations | 第10-12页 |
| Abstracts | 第12-13页 |
| 中文摘要 | 第14-16页 |
| CHAPTER ONE INTRODUCTION AND LITERATURE REVIEW | 第16-29页 |
| Introduction | 第16-18页 |
| 1. The effects of CCs and their apoptosis on oocyte quality during IVM | 第18-23页 |
| 1.1 The effects of CCs on oocyte quality during IVM | 第18-20页 |
| 1.2 The effects of CCs apoptosis on oocyte maturation and embryo development | 第20-22页 |
| 1.3 The effects of OSFs on CCs apoptosis during IVM | 第22-23页 |
| 2. The effects and signaling pathway of BMP15 and GDF9 | 第23-29页 |
| 2.1 The biological effects of BMP15 and GDF9 | 第24-25页 |
| 2.2 Signaling pathway of BMP15 and GDF9 during IVM | 第25-29页 |
| CHAPTER TWO EXPERIMENTS AND RESULTS EXPERIMENTAL PURPOSE | 第29-62页 |
| EXPERIMENT ONE BMP15 and GDF9 m RNA expression in bovine oocyte during IVM | 第30-42页 |
| 1. Introduction | 第30页 |
| 2. Materials and Methods | 第30-35页 |
| 2.1 Chemicals and media | 第31页 |
| 2.2 Instruments and equipments | 第31页 |
| 2.3 Experimental methods | 第31-35页 |
| 2.3.1 Experiment design and technical route | 第31-32页 |
| 2.3.2 In vitro maturation | 第32-33页 |
| 2.3.3 Nuclear staining of GV oocytes | 第33页 |
| 2.3.4 Sampling denuded oocytes at different time after IVM | 第33页 |
| 2.3.5 RNA extraction and cDNA synthesis | 第33-34页 |
| 2.3.6 Real-time quantitative PCR for BMP15 and GDF9 | 第34页 |
| 2.3.7 Statistical analysis | 第34-35页 |
| 3. Results | 第35-39页 |
| 3.1 GV stage observation of oocyte before IVM | 第35-36页 |
| 3.2 The results of BMP15 m RNA expression in bovine oocytes by qRT-PCR | 第36-37页 |
| 3.2.1 The solubility curve of BMP15 mRNA | 第36页 |
| 3.2.2 BMP15 mRNA standard curve | 第36-37页 |
| 3.2.3 BMP15 mRNA expression in bovine oocyte during IVM | 第37页 |
| 3.3 The results of GDF9 m RNA expression in bovine oocytes by qRT-PCR | 第37-39页 |
| 3.3.1 The solubility curve of GDF9 mRNA | 第37-38页 |
| 3.3.2 The standard curve of GDF9 mRNA | 第38页 |
| 3.3.3 The expression of GDF9 mRNA in bovine oocyte during IVM | 第38-39页 |
| 4. Discussion | 第39-41页 |
| 5. Conclusion | 第41-42页 |
| EXPERIMENT TWO Effects of BMP15 and GDF9 on apoptosis of bovine CCs during IVM | 第42-54页 |
| 1. Introduction | 第42页 |
| 2. Materials and Methods | 第42-47页 |
| 2.1 Chemicals and media | 第42-43页 |
| 2.2 Instrument and equipment | 第43页 |
| 2.3 Experimental methods | 第43-47页 |
| 2.3.1 Experiment design and Technical route | 第43-44页 |
| 2.3.2 In vitro fertilization | 第44页 |
| 2.3.3 In vitro embryo culture | 第44-45页 |
| 2.3.4 The reconstitution and the addition of BMP15 and GDF9 | 第45页 |
| 2.3.5 Sampling of CCs from collected COCs | 第45页 |
| 2.3.6 Real-time quantitative PCR for anti-apoptotic and pro-apoptotic genes | 第45-46页 |
| 2.3.7 Western blot analysis | 第46-47页 |
| 2.3.8 Statistical analysis | 第47页 |
| 3. Results | 第47-50页 |
| 3.1 The rate and quality of blastocyst after bovine IVF with GDF9 addition | 第47-48页 |
| 3.2 The mRNA expression anti-apoptotic genes and pro-apoptotic genes in CC withBMP15 or GDF9 addition | 第48-50页 |
| 3.3 The protein expression level of Bim and P53 in CC with BMP15 or GDF9 addition | 第50页 |
| 4. Discussion | 第50-52页 |
| 5. Conclusion | 第52-54页 |
| EXPERIMENT THREE The signaling pathway of GDF9 on the apoptosis of bovine CCs during IVM | 第54-62页 |
| 1. Introduction | 第54页 |
| 2. Materials and Methods | 第54-56页 |
| 2.1 Chemicals and media | 第55页 |
| 2.2 Instrument and equipment | 第55页 |
| 2.3 Experimental methods | 第55-56页 |
| 2.3.1 Technical route and experimental design | 第55-56页 |
| 2.3.2 Treatment with various inhibitors | 第56页 |
| 2.3.3 Western blot analysis | 第56页 |
| 2.3.4 Statistical analysis | 第56页 |
| 3. Results | 第56-57页 |
| 3.1 The signaling pathway of GDF9 on the apoptosis of the bovine CCs during IVM | 第56-57页 |
| 4. Discussion | 第57-60页 |
| 5. Conclusion | 第60-62页 |
| CHAPTER THREE CONCLUSIONS | 第62-64页 |
| Reference | 第64-72页 |
| ACKNOWLEDGEMENTS | 第72-74页 |
| Resume | 第74页 |
