声明
Contents
摘要
Abstract
Abbreviations
Chapter 1
1 Introduction
1．1 Rationale
1．2 Hypothesis and aims
Chapter 2
2．Review of literature
2．1 Overview of TGF β Super-family and signaling pathway
2．2 Biology of Follicle development
2．3 Hormonal control on folliculogenesis
2．4．Oocyte granulose cell interaction
2．5 Transgenic RNAi in mouse Oocytes
2．6 Inhibins and premature failure
2．7 Physiological consequence of the inhibin mutation in ovary
2．8 Overview of RNA interference
2．9 Apoptosis and cell cycle regulation
Chapter 3
3 Materials and Methods
3．1 Construction ofrecombinant vectors and transgenic mice
3．2 Mieroinjection and Transgenic mice production
3．3 Experimental animal and ethics statement
3．4 Animal breeding and genotyping
3．5 Immunohistochemistry
3．6 Isolation and culture of primary GCs
3．7 RNA extraction and reverse transcription polymerase chain reaction(RT-PCR)
3．8 Quantification of gene expression by quantitative polymerase chain reaction(qPCR)
3．9 Western blotting
3．10 Detection of cell apoptosis
3．11 Cell cycle analysis
3．12 Hormonal assays
3．13 Fertility measurement
3．14 Determination of onset of puberty and estrus cycle
3．15 Super-ovulation and oocyte count procedure
Chapter 4
4 Results
4．1 INH α-subunit gene expression in GCs
4．2 The RNAi-M induced silencing effect in GCs
4．3 Silencing of INH α altered the expression level of INHβB，bcl2，Caspase3，Kitl，GDF9 and TGFβⅢ in GCs
4．4 Silencing of INH α induced mice GCs apoptosis
4．5 Knockdown of INH α altered the growth and proliferation of the mouse GCS
4．6 Knockdown of INH α altered the hormone secretions of INF α and FSH
4．7 Reduced the fertility in RNAi INH α mice
4．8 Delayed onset of puberty and estrus cycle
Chapter 5
5 Discussion，Conclusion and future directions
5．1 Discussion
5．2 Conclusions
5．3 Future directions
References
Acknowledgement
Dedication
Award and publications
ANNEX
