| 摘要 | 第10-12页 |
| Abstract | 第12-13页 |
| Abbreviations | 第14-15页 |
| Chapter 1 Introduction | 第15-18页 |
| 1.1 Rationale | 第16-17页 |
| 1.2 Hypothesis and aims | 第17-18页 |
| Chapter 2 Review of literature | 第18-46页 |
| 2.1 Overview of TGF β Super-family and signaling pathway | 第18-26页 |
| 2.1.1 Ligands of TGF β Super-family | 第18-19页 |
| 2.1.2 Inhibins | 第19-20页 |
| 2.1.3 TGFβ superfamily signaling receptors | 第20-22页 |
| 2.1.4 Inhibin receptors | 第22页 |
| 2.1.5 TGFβ Super family signaling | 第22-24页 |
| 2.1.6 Inhibin Signaling | 第24-25页 |
| 2.1.7 Intraovarian role of inhibins | 第25-26页 |
| 2.1.8 The roles of Inhibins in folliculogenesis | 第26页 |
| 2.2 Biology of Follicle development | 第26-33页 |
| 2.2.1 Overview of ovarian Folliculogenesis | 第26-28页 |
| 2.2.2 Primordial germ cell formation and migration | 第28-30页 |
| 2.2.3 Early antral follicles development | 第30-31页 |
| 2.2.4 Antral follicle growth | 第31-33页 |
| 2.2.5 Ovulation and CL formation | 第33页 |
| 2.3 Hormonal control on folliculogenesis | 第33-34页 |
| 2.4. Oocyte granulose cell interaction | 第34-37页 |
| 2.4.1 Oocyte control granulosa and cumulus cell function | 第34-35页 |
| 2.4.2 Follicular control of oocyte maturation | 第35-37页 |
| 2.5 Transgenic RNAi in mouse Oocytes | 第37-38页 |
| 2.6 Inhibins and premature failure | 第38-39页 |
| 2.7 Physiological consequence of the inhibin mutation in ovary | 第39-40页 |
| 2.8 Overview of RNA interference | 第40-43页 |
| 2.8.1 Mechanisms of gene silencing by siRNA | 第41-42页 |
| 2.8.2 Mechanisms of gene silencing by shRNA | 第42-43页 |
| 2.9 Apoptosis and cell cycle regulation | 第43-46页 |
| Chapter 3 Materials and Methods | 第46-59页 |
| 3.1 Construction of recombinant vectors and transgenic mice | 第46-47页 |
| 3.2 Microinjection and Transgenic mice production | 第47-51页 |
| 3.2.1 Purification of transgene DNA fragment | 第47-48页 |
| 3.2.2 Super-ovulation procedure | 第48页 |
| 3.2.3 Production of Pseudo pregnant recipient females | 第48页 |
| 3.2.4 Collection of embryos | 第48-49页 |
| 3.2.5 Preparation of transgene DNA for Microinjection | 第49页 |
| 3.2.6 Microinjection | 第49-50页 |
| 3.2.7 Transfer of embryos to recipient females | 第50-51页 |
| 3.3 Experimental animal and ethics statement | 第51-52页 |
| 3.4 Animal breeding and genotyping | 第52-53页 |
| 3.5 Immunohistochemistry | 第53-54页 |
| 3.6 Isolation and culture of primary GCs | 第54-55页 |
| 3.7 RNA extraction and reverse transcription polymerase chain reaction(RT-PCR) | 第55页 |
| 3.8 Quantification of gene expression by quantitative polymerase chain reaction(qPCR) | 第55页 |
| 3.9 Western blotting | 第55-56页 |
| 3.10 Detection of cell apoptosis | 第56页 |
| 3.11 Cell cycle analysis | 第56-57页 |
| 3.12 Hormonal assays | 第57页 |
| 3.13 Fertility measurement | 第57页 |
| 3.14 Determination of onset of puberty and estrus cycle | 第57-58页 |
| 3.15 Super-ovulation and oocyte count procedure | 第58-59页 |
| Chapter 4 Results | 第59-66页 |
| 4.1 INHα-subunit gene expression in GCs | 第59页 |
| 4.2 The RNAi-M induced silencing effect in GCs | 第59-60页 |
| 4.3 Silencing of INH α altered the expression level of INHβB,bcl2,Caspase3,Kitl, GDF9 and TGFβⅢ in GCs | 第60-61页 |
| 4.4 Silencing of INH α induced mice GCs apoptosis | 第61-62页 |
| 4.5 Knockdown of INHα altered the growth and proliferation of the mouse GCs | 第62页 |
| 4.6 Knockdown of INHα altered the hormone secretions of INH α and FSH | 第62-63页 |
| 4.7 Reduced the fertility in RNAi INH α mice | 第63-64页 |
| 4.8 Delayed onset of puberty and estrus cycle | 第64-66页 |
| Chapter 5 Discussion,Conclusion and future directions | 第66-73页 |
| 5.1 Discussion | 第66-72页 |
| 5.2 Conclusions | 第72页 |
| 5.3 Future directions | 第72-73页 |
| References | 第73-88页 |
| Acknowledgement | 第88-89页 |
| Dedication | 第89-90页 |
| Award and publications | 第90-91页 |
| ANNEX | 第91-93页 |
