RNA Interference as a tool to study the function of MAD2 in mouse oocyte meiotic maturation.
Spindle checkpoint proteins control entry into anaphase and chromosome segregation. As a member of spindle checkpoint proteins, MAD2 takes a central role in the regulation of anaphase onset and genome integrity. Here, we used MAD2 siRNA transfection approach to study MAD2 functions during mouse oocyte meiotic maturation in vitro. Real-time PCR and laser scanning confocal microscopy showed that we successfully downregulated MAD2 transcript and protein expression. We further demonstrated that MAD2 downregulation resulted in a shortened duration of meiosis I and meiotic spindle abnormality, suggesting the function of MAD2 in mouse oocyte meiotic maturation. We also showed that MAD2 interference to some extent decreased GVBD rate, but increased apoptosis in mouse oocytes. In conclusion, our study shows that siRNA transfection is an effective tool to study MAD2 functions, and our results provide further evidence for the role of MAD2 as a spindle checkpoint protein in mouse oocytes.
Wang
Jian-Ying
JY
Key Laboratory of Animal Resistance Research, College of Life Science, Shandong Normal University, Jinan, PR China.
Lei
Zi-Li
ZL
Nan
Chang-Long
CL
Yin
Shen
S
Liu
Jinghe
J
Hou
Yi
Y
Li
Yun-Long
YL
Chen
Da-Yuan
DY
Sun
Qing-Yuan
QY
D000818:Animals; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D005260:Female; D064246:Mad2 Proteins; D008540:Meiosis / Q000187:drug effects / Q000235:genetics; D049469:Meiotic Prophase I / Q000187:drug effects / Q000235:genetics*; D051379:Mice; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development*; D009866:Oogenesis / Q000187:drug effects / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology; D008941:Spindle Apparatus / Q000187:drug effects; D014162:Transfection
D018797:Cell Cycle Proteins; D064246:Mad2 Proteins; C577698:Mad2l1 protein, mouse; D034741:RNA, Small Interfering