Identification and characterization of an oocyte factor required for porcine nuclear reprogramming.
Nuclear reprogramming of somatic cells can be induced by oocyte factors. Despite numerous attempts, the factors responsible for successful nuclear reprogramming remain elusive. In the present study, we found that porcine oocytes with the first polar body collected at 42 h of in vitro maturation had a stronger ability to support early development of cloned embryos than porcine oocytes with the first polar body collected at 33 h of in vitro maturation. To explore the key reprogramming factors responsible for the difference, we compared proteome signatures of the two groups of oocytes. 18 differentially expressed proteins between these two groups of oocytes were discovered by mass spectrometry (MS). Among these proteins, we especially focused on vimentin (VIM). A certain amount of VIM protein was stored in oocytes and accumulated during oocyte maturation, and maternal VIM was specifically incorporated into transferred somatic nuclei during nuclear reprogramming. When maternal VIM function was inhibited by anti-VIM antibody, the rate of cloned embryos developing to blastocysts was significantly lower than that of IgG antibody-injected embryos and non-injected embryos (12.24 versus 22.57 and 21.10%; p < 0.05), but the development of in vitro fertilization and parthenogenetic activation embryos was not affected. Furthermore, we found that DNA double strand breaks dramatically increased and that the p53 pathway was activated in cloned embryos when VIM function was inhibited. This study demonstrates that maternal VIM, as a genomic protector, is crucial for nuclear reprogramming in porcine cloned embryos.
Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Qingran
Q
Kong
Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Bingteng
B
Xie
Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Jingyu
J
Li
Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Yanjun
Y
Huan
Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Tianqing
T
Huang
Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Renyue
R
Wei
Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Jiawei
J
Lv
Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Shichao
S
Liu
Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin 150030, China. Electronic address: liu086@yahoo.com.
Zhonghua
Z
Liu
D000818:Animals; D001755:Blastocyst / Q000378:metabolism / Q000502:physiology; D065150:Cellular Reprogramming; D019976:Cloning, Organism; D047108:Embryonic Development; D005260:Female; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000378:metabolism / Q000502:physiology*; D059705:Polar Bodies / Q000378:metabolism / Q000502:physiology; D013552:Swine / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D014746:Vimentin / Q000037:antagonists & inhibitors / Q000378:metabolism*
D014746:Vimentin
Chromatin Remodeling; Cloning; DNA Damage; Oocyte; Pig; Reprogramming; Vimentin