| 摘要 | 第12-14页 |
| ABSTRACT | 第14-16页 |
| LIST OF ABBERIVATIONS | 第17-20页 |
| CHAPTER-1: LITERATURE REVIEW | 第20-41页 |
| 1.1. Folliculogenesis | 第20-25页 |
| 1.1.1. Pre-antral folliculogenesis | 第22-23页 |
| 1.1.2. Antral folliculogenesis | 第23-25页 |
| 1.2. Follicular Atresia | 第25-27页 |
| 1.2.1. Apoptosis a tool for follicular atresia | 第25-27页 |
| 1.3. Influence of Oocyte Factor in Folliculogenesis | 第27-29页 |
| 1.4. Ephrins and Eph Receptors Family | 第29-39页 |
| 1.4.1. Eph receptors gene | 第31-32页 |
| 1.4.2. Ephrin gene | 第32-33页 |
| 1.4.3. Novel mechanism of actions of ephrin-Eph signaling | 第33-36页 |
| 1.4.3.1. Ephrin-Eph signalling (forward signaling) | 第34-35页 |
| 1.4.3.2. Eph-ephrin signalling (reverse signaling) | 第35-36页 |
| 1.4.4. Ephrin-Eph signaling in cellular apoptosis and proliferation | 第36-39页 |
| 1.5. General Objectives of the Research | 第39-41页 |
| CHAPTER 2: REGULATORY ROLE AND NOVEL SIGNALING PATHWAY OF EFNA5 IN MOUSE PRIMARY GRANULOSA CELLS APOPTOSIS ANDPROLIFERATION | 第41-65页 |
| 2.0 INTRODUCTION | 第41-43页 |
| 2.1. Hypothesis of the Study | 第43页 |
| 2.2. Objectives of the Study | 第43-44页 |
| 2.3. Materials and Methods | 第44-50页 |
| 2.3.1 Management of experimental animals | 第44页 |
| 2.3.2. Mouse GC isolation and culture | 第44-45页 |
| 2.3.3. Immunohistochemistry (IHC) analysis | 第45页 |
| 2.3.4. Ephrin A5 si RNA (si Efna5) transfection | 第45-46页 |
| 2.3.5. RNA extraction and reverse-transcription polymerase chain reaction (RT-PCR) | 第46页 |
| 2.3.6. Quantitative Real-Time polymerase chain reaction (q RT-PCR) | 第46-47页 |
| 2.3.7. Protein extraction and western blotting | 第47-48页 |
| 2.3.8. Cell apoptosis detection | 第48-49页 |
| 2.3.9. Cell proliferation assay | 第49页 |
| 2.3.10. Cell cycle assay | 第49页 |
| 2.3.11. Enzyme-linked immunosorbent assay for steroid hormones (ELISA) | 第49-50页 |
| 2.3.12. Statistical analysis | 第50页 |
| 2.4. Results | 第50-60页 |
| 2.4.1. Expression of Efna5 in mouse ovarian tissues | 第50-51页 |
| 2.4.2. Efna5 and cognate receptors are developmentally regulated in mouse GC | 第51-52页 |
| 2.4.3. Efna5 expression is efficiently downregulated by si Efna5 | 第52页 |
| 2.4.4. m RNA expression levels of Efna5 receptors are inhibited by si Efna5 | 第52-53页 |
| 2.4.5. Silencing of Efna5 suppresses mouse GC apoptosis in vitro | 第53-54页 |
| 2.4.6. Downregulation of Efna5 dysregulated the expression of apoptotic regulators in mouse GC in vitro | 第54-56页 |
| 2.4.7. Silencing of Efna5 promotes mouse GC proliferation in vitro without affecting cell cycle progression | 第56-57页 |
| 2.4.8. Inhibition of Efna5 regulates mouse GC proliferation by activating the Erk1/2 and Akt phosphorylation pathways | 第57页 |
| 2.4.9.Knockdown of Efna5 alters the concentration of steroidogenesis and expression of oocyte-secreted factors in mouse GC in vitro | 第57-59页 |
| 2.4.10. Downregulation of Efna5 inhibited m RNA expression of VEGFA in mouse GC | 第59-60页 |
| 2.5. Discussion | 第60-65页 |
| CHAPTER-3:TRANSCRIPTOME PROFILING OF EPHRINA5 si RNA TREATEDMOUSE GRANULOSA CELLS | 第65-81页 |
| 3.0 INTRODUCTION | 第65-66页 |
| 3.1. Why Gene Expressions Profiling by RNA-Seq? | 第66页 |
| 3.2. Objectives | 第66页 |
| 3.3. Materials and Methods | 第66-70页 |
| 3.3.1. Experimental animal management | 第66-67页 |
| 3.3.2. Granulosa cells isolation and culturing | 第67页 |
| 3.3.3. Ephrin A5 si RNA (si Efna5) transfection | 第67-68页 |
| 3.3.4. Total RNA extraction and library preparations | 第68页 |
| 3.3.5. Assembling and mapping of fragments | 第68-69页 |
| 3.3.6. Analysis of RNA-seq data | 第69页 |
| 3.3.7. Alternative splicing, Idel and SNP analysis | 第69页 |
| 3.3.8. KEGG pathway and enrichment analyses | 第69-70页 |
| 3.3.9. Transcriptome profiling analyses work flow | 第70页 |
| 3.4. Result and Discussion | 第70-81页 |
| 3.4.1. Analysis of RNA-seq reads mapping and quality control | 第70-71页 |
| 3.4.2. Samples expression density analysis | 第71-72页 |
| 3.4.3. RNA-seq correlation and principal component analysis | 第72-73页 |
| 3.4.4. SNP, In Del, Alternative splicing analysis | 第73页 |
| 3.4.5.Alternative splicing analysis | 第73-74页 |
| 3.4.6. Heatmap analysis of expressed genes | 第74-75页 |
| 3.4.7. Differentially expressed genes (DEGs) | 第75-77页 |
| 3.4.8 Gene ontology (GO) enrichment analysis | 第77-79页 |
| 3.4.9. Pathway and network enrichment analysis | 第79-81页 |
| CHAPTER-4: COMPREHENSIVE STRUCTURAL AND EPIGENETIC ANALYSIS OFMOUSE EPHRINA5 AND ITS RECEPTORS PROTEINS | 第81-119页 |
| 4.0 INTRODUCTION | 第81-83页 |
| 4.1. Objectives of the Study | 第83页 |
| 4.2. Methodologies | 第83-87页 |
| 4.2.1. Exploration and mining of mouse Efna5 and its receptor structure and sequences | 第83-84页 |
| 4.2.2. 3D structure prediction and validation of mouse Efna5 and its receptors | 第84页 |
| 4.2.3 Disordered analysis of mouse Efna5 and its receptor proteins | 第84-85页 |
| 4.2.4. Analysis of mouse Efna5 and its receptor protein ligands and domain | 第85-86页 |
| 4.2.5. Analysis of cross-correlations and distance fluctuations of mouse Efna5 and its receptorproteins | 第86页 |
| 4.2.6. Protein interactions and pathway analysis | 第86页 |
| 4.2.7 Predicting consensus sequence alignment and secondary structure | 第86-87页 |
| 4.2.8. Cp G island prediction | 第87页 |
| 4.3 Result | 第87-113页 |
| 4.3.1. Mouse Efna5 and its receptors 3D protein modeling | 第87-88页 |
| 4.3.2. 3D protein modeling validation | 第88-90页 |
| 4.3.2.1. Pro SA and VERFIY based validation of the predicted protein structure | 第88-89页 |
| 4.3.2.2. Physicochemical based validation of the predicted structure | 第89-90页 |
| 4.3.3. Disordered mouse Efna5 and its receptors proteins structure | 第90-92页 |
| 4.3.4 Predicted ligand binding site of mouse Efna5 and Epha5 proteins | 第92页 |
| 4.3.5 Predicted ligand binding site of mouse Epha3 and Ephb2 proteins | 第92-93页 |
| 4.3.6. Analysis of predited ligand binding sites of mouse Efna5 and receptor proteins | 第93-94页 |
| 4.3.7. Mouse Efna5 and its receptors ligand consensus module | 第94-95页 |
| 4.3.8. Amino acids composition interacting ligands of mouse Efan5 and its receptor proteins | 第95-96页 |
| 4.3.9 Prediction of FTSite for mouse Efna5 and its receptor proteins | 第96-97页 |
| 4.3.10 Prediction of FTSite of mouse Ephb2 protein | 第97-98页 |
| 4.3.11 Domain structure of mouse Efna5 and its receptors proteins | 第98-99页 |
| 4.3.12. Cross-correlation analysis of mouse Efna5 and its receptors proteins | 第99-100页 |
| 4.3.13 Predicted signal communication analysis of mouse Efna5 and Epha5 proteins | 第100-101页 |
| 4.3.14 Distance fluctuation analysis of Efna5 and its receptor proteins | 第101-102页 |
| 4.3.15 Ramchandran plot of mouse Efna5 and its receptors proteins | 第102-103页 |
| 4.3.16. Protein interaction analysis of mouse Efna5 and its receptors Protein | 第103-105页 |
| 4.3.16.1. Predicted protein-protein interaction action view | 第103-104页 |
| 4.3.16.2 Predicted protein-protein interaction evidence view | 第104-105页 |
| 4.3.17 Pathway analysis of mouse Efna5 proteins | 第105-106页 |
| 4.3.18. Pathway analysis of mouse Eph receptors proteins | 第106-107页 |
| 4.3.19. Predicted consensus sequence alignment of mouse Efna5 and Ephb2 proteins | 第107-108页 |
| 4.3.20. Predicted consensus structural alignment of mouse Efna5 and Ephb2 proteins | 第108-109页 |
| 4.3.21. Predicted secondary structure proteins | 第109-112页 |
| 4.4.21.1. Predicted secondary structure of mouse Efna5 protein | 第109-110页 |
| 4.3.21.2. Predicted secondary structure of mouse Epha5 proteins | 第110-111页 |
| 4.3.21.3. Predicted secondary structure of mouse Ephb2 proteins | 第111-112页 |
| 4.3.22. CpG island prediction of mouse Efna5 and its downstream receptors | 第112-113页 |
| 4.4. Discussions | 第113-119页 |
| CHAPTER-5: CONCLUSIONS AND FUTURE DIRECTIONS | 第119-123页 |
| 5.1. Study-Ⅰ Conclusion | 第119-120页 |
| 5.2. Study-Ⅱ Conclusion | 第120-121页 |
| 5.3. Study-Ⅲ Conclusion | 第121-122页 |
| 5.4. Future Directions | 第122-123页 |
| 6. REFERENCES | 第123-153页 |
| 7. APPENDCES | 第153-172页 |
| 8. ACKNOWLEDGEMENTS | 第172-173页 |
| 9. DEDICATION | 第173-174页 |
| 10. LIST OF PUBLICATIONS | 第174-176页 |
| 作者简介 | 第176-178页 |
