원문정보
초록
영어
Cell adhesion plays an important role in the differentiation of the morphogenesis and the trophectoderm epithelium of the blastocyst. In the porcine embryo, CDH1 mediated adhesion initiates at compaction before blastocyst formation, regulated post-translationally via protein kinase C and other signaling molecules. Here we focus on muscle RAS oncogene homolog (M-RAS), which is the closest relative to the RAS related proteins and shares most regulatory and effector interactions. To characterize the effects of M-RAS on embryo compaction, we used gain- and loss-of-function strategies in porcine embryos, in which M-RAS gene structure and protein sequence are conserved. We showed that knockdown of M-RAS in zygotes reduced embryo development abilities and CDH1 expression. Moreover, the phosphorylation of ERK was also decreased in M-RAS KD embryos. Overexpression of M-RAS allows M-RAS KD embryos to rescue the embryo compaction and blastocyst formation. Collectively, these results highlight novel conserved and multiple effects of M-RAS during porcine embryo development.
목차
INTRODUCTION
MATERIALS AND METHODS
Reagents
Oocyte collection and in vitro maturation (IVM)
Parthenogenetic activation and in vitro culture (IVC)
M-RAS double-stranded RNA preparation
Cloning and in vitro mRNA synthesis
Microinjection
Immunofluorescence and confocal microscopy
Real-time RT-PCR
Western blot analysis
Statistical analysis
RESULTS
Expression patterns of M-RAS during porcine embryo development
Effects of M-RAS knockdown on early porcine embryonic development
The active M-RAS regulates cell-cell adhesion by modulating the E-cadherin which correlates with ERK cascade activation
Overexpression of M-RAS rescues the disrupt of embryonic compaction after M-RAS knockdown
DISCUSSION
CONFLICTS OF INTEREST
ACKNOWLEDGEMENTS
AUTHOR CONTRIBUTIONS
AUTHOR’S POSITION AND ORCID NO.
REFERENCES