声明
LIST ACRONYMS
GENERAL INTRODUCTION
CHAPTER ONE LITERATURE REVIEW
1.1 BACKGROUND
1.2 THE REPRODUCTIVE TRACT
1.2.1 Ovary
1.2.2 Primordial and primary follicles
1.2.3 Prehierarchical follicles
1.3 HEN OVARIAN FOLLICLE CELLS
1.3.1 Oocyte
1.3.2 Granulosa cells
1.3.3 Theca cells
1.4 AVIAN FOLLICLE SELECTION AND FORMATION OF THE PRE-OVULATORY ORDER
1.5 FOLLICLE ATRESIA
1.6 VESTIGIAL LIKE (VGLL) GENE FAMILY
1.6.1 Chicken VGLL1
1.6.2 Chicken VGLL4
1.7 RESEARCH OBJECTIVES
1.7.1 Overall object
1.7.2 Specific objectives
1.8 THESIS ORGANIZATION
CHAPTER 2 IN SILICO ANALYSIS OF VGLL1 AND VGLL4
2.1 INTRODUCTION
2.2 METHODS
2.2.1 Retrieval and analysis of VGLL4 protein, Gene sequence and Coding sequences
2.2.2 Phylogenetic Analysis
2.2.3 Physiochemical properties of the VGLL 1 and VGLL4 protein
2.2.4 Exon structural analysis
2.2.5 Conserved motif analysis
2.2.6 Conserved synteny analysis
2.2.7 Analysis of protein-protein interactions
2.3 RESULTS
2.3.1 In silico analysis of VGLL1
2.3.2 In silico analysis of VGLL4
2.4 DISCUSSION
CHAPTER3 3 EXPRESSION OF VGLL1 AND VGLL4 IN GRANULOSA CELLS OF HEN OVARIAN PREHIERARCHICAL FOLLICLES
3.1 INTRODUCTION
3.2 METHODS
3.2.1 Chickens and sampling
3.2.2 Harvesting follicles
3.2.3 Extraction of RNA
3.2.4 Reverse transcription
3.2.5 Primer Design
3.2.6 Real-time quantitative RT-PCR
3.2.7 Statistical analysis
3.3 RESULTS
3.3.1 Relative expression levels of VGLL1 ovarian Granulosa Cells by Real time PCR
3.3.2 Relative expression levels of VGLL4 ovarian Granulosa Cells by Real time PCR
3.4 DISCUSSION
CHAPTER 4 Effect of knocking down VGLL1 and VGLL4 genes on granulosa cell Apoptosis, Proliferation and Steroidogenesis of hen ovarian prehierarchical follicles
4.1 INTRODUCTION
4.2 MATERIALS AND METHODS
4.2.1 Chickens and sampling
4.2.1 Real-time quantitative RT-PCR
4.2.2 Culture of granulosa cells and cell transfection of VGLL1 expression
4.2.3 Flow Cytometry detection of apoptosis
4.2.4 Cell proliferation assay
4.2.5 Data analysis
4.3 RESULTS
4.3.1 SELECTION OFAPPROPRIATE SMALL INTERFERENCE RNA (siRNA)
4.3.2 EFFECTS OF SILENCING TARGET GENES (VGLL1 AND VGLL4) ON APOPTOSIS
4.3.3 EFFECTS OF KNOCKDOWN OF TARGET GENES ON GRANULOSA CELL PROLIFERATION
4.3.4 EFFECTS OF SILENCING TARGET GENES (VGLL1 AND VGLL4) ON STEROIDOGENESIS
4.4 DISCUSSION
CHAPTER 5 CONCLUSION
参考文献
APPENDICES
AUTHOR’S INFORMATION
致谢
吉林农业大学;
