돼지와 생쥐 난자의 체외성숙과 배아의 발달에 영향을 미치는 요인들
: Factors Affecting in vitro Oocyte Maturation and Early Embryo Development in Pigs and Mice
초록 (Abstract)- We investigated the function of genes (GDF9, BMP15 and JMY) in porcine and mouse oocytes maturation, activation and embryos development in vitro. Chapter II: Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) are member...
- We investigated the function of genes (GDF9, BMP15 and JMY) in porcine and mouse oocytes maturation, activation and embryos development in vitro.
Chapter II: Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) are members of the transforming growth factor-b (TGF-β) family, and their roles in oocyte maturation and cumulus expansion are well known in the mouse and0 human, but not in the pig. We investigated GDF9 and BMP15 expressions in porcine oocytes during in vitro maturation. A significant increase in the mRNA levels of GDF9 and BMP15 was observed at germinal vesicle break- down, with expression levels peaking at metaphase I (MI), but decreasing at metaphase II (MII). GDF9 and BMP15 protein localized to the oocyte cytoplasm. While treatment with GDF9 and BMP15 increased the expression of genes involved in both oocyte maturation (c-mos, cyclinb1 and cdc2) and cumulus expansion (has2, ptgs2, ptx3 and tnfaip6), SB431542 (a TGFβ–GDF9 inhibitor) decreased meiotic maturation at MII. Following parthenogenetic activation, the percentage of blastocysts in SB431542 treatment was lower than in the control (41.3% and 74.4%, respectively). Treatment with GDF9 and BMP15 also increased the mRNA levels of maternal genes such as c-mos (a regulatory subunit of mitogen-activated protein kinase (MAPK)), and cyclinb1 and cdc2 (regulatory subunits of maturation/ M-phase-promoting factor (MPF)); however, SB431542 significantly decreased their mRNA levels. These data were supported by poly (A) - test PCR and protein activity analyses. Our results show that GDF9 and BMP15 participate in cumulus expansion and that they stimulate MPF and MAPK activities in porcine oocytes during in vitro maturation.
목차 (Table of Contents)- CHAPTER I. LITERATURE REVIEW 1
- 1. OOCYTE MEIOTIC MATURATION 1
- 2. MECHANISM OF MPF REGULATE MEIOTIC DIVISION 1
- 3. PREIMPLANTATION EMBRYONIC DEVELOPMENT 2
- 4. REFERENCE 4
- CHAPTER II. EFFECTS OF GROWTH DIFFERENTIATION FACTOR 9 AND BONE MORPHOGENETIC PROTEIN 15 ON THE IN VITRO MATURATION OF PORCINE OOCYTES. 6
- ABSTRACT 6
- INTRODUCTION 7
- MATERIALS AND METHODS 10
- RESULTS 15
- DISCUSSION 19
- REFERENCES 30
- CHAPTER III. JMY FUNCTIONS AS ACTIN NUCLEATION- PROMOTING FACTOR AND MEDIATOR FOR P53-MEDIATED DNA DAMAGE IN PORCINE OOCYTES 37
- ABSTRACT 37
- INTRODUCTION 38
- MATERIALS AND METHODS 39
- RESULTS 45
- DISCUSSION 56
- REFERENCES 58
- CHAPTER IV. JMY IS ESSENTIAL FOR PORCINE EARLY EMBRYO DEVELOPMENT 63
- ABSTRACT 63
- INTRODUCTION 64
- MATERIALS AND METHODS 65
- RESULTS 71
- DISCUSSION 79
- REFERENCES 81
- CHAPTER V. DUAL ROLE OF JMY IN MOUSE PREIMPLANTATION EMBRYOS 85
- ABSTRACT 85
- INTRODUCTION 86
- MATERIALS AND METHODS 88
- RESULTS 93
- DISCUSSION 107
- REFERENCES 111
- CHAPTER VII 118
- GENERAL SUMMARY 118
- ACKNOWLEDGEMENTS 120