Survivin acts as anti-apoptotic factor during the development of bovine pre-implantation embryos.
Survivin, an inhibitor of apoptotic protein containing a single baculoviral inhibit apoptotic protein repeat domain, is a bifunctional protein that suppresses apoptosis and regulates cell division. Thus, we used double stranded RNA (dsRNA) interference to manipulate survivin expression in bovine embryos and analyze its role in blocking apoptosis and facilitating development of pre-implantation embryos. In vitro fertilized embryos (1-cell) were injected with survivin dsRNA, and expression of survivin mRNA was evaluated by real-time quantitative RT-PCR. To analyze survivin protein expression, we performed immunocytochemistry using a rabbit anti-bovine suvivin antibody. Expression levels of survivin mRNA and protein were decreased in the dsRNA group compared to the sham group. Rates of in vitro blastocyst development were lower in the survivin dsRNA-injected group than in the sham-injected group. Also, the total cell number seen in blastocysts was decreased in the dsRNA group. TUNEL assays of DNA fragmentation indicated an increased apoptotic index in the dsRNA group compared to the sham group. These results indicate that survivin is important for optimal development of bovine blastocysts and confirm that survivin expression suppresses apoptosis of pre-implantation embryos.
Park
Sae Young
SY
Maria Infertility Hospital Medical Institute/Maria Biotech, Seoul, and Chungbuk National University, Chungbuk, South Korea. spark@cheju.ac.kr
Kim
Eun Young
EY
Jeon
Kilsoo
K
Cui
Xiang-Shun
XS
Lee
Won Don
WD
Kim
Nam-Hyung
NH
Park
Se Pill
SP
Lim
Jin Ho
JH
D000818:Animals; D017209:Apoptosis; D001755:Blastocyst / Q000378:metabolism / Q000502:physiology*; D053148:Caspase 3 / Q000096:biosynthesis; D002417:Cattle; D002478:Cells, Cultured; D046149:Embryo Culture Techniques; D005260:Female; D020869:Gene Expression Profiling; D008297:Male; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis* / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D008869:Microtubule-Associated Proteins; D012333:RNA, Messenger; D053148:Caspase 3