The cytoplasm of mouse germinal vesicle stage oocytes can enhance somatic cell nuclear reprogramming.
In mammalian cloning, evidence suggests that genomic reprogramming factors are located in the nucleus rather than the cytoplasm of oocytes or zygotes. However, little is known about the mechanisms of reprogramming, and new methods using nuclear factors have not succeeded in producing cloned mice from differentiated somatic cell nuclei. We aimed to determine whether there are functional reprogramming factors present in the cytoplasm of germinal vesicle stage (GV) oocytes. We found that the GV oocyte cytoplasm could remodel somatic cell nuclei, completely demethylate histone H3 at lysine 9 and partially deacetylate histone H3 at lysines 9 and 14. Moreover, cytoplasmic lysates of GV oocytes promoted somatic cell reprogramming and cloned embryo development, when assessed by measuring histone H3-K9 hypomethylation, Oct4 and Cdx2 expression in blastocysts, and the production of cloned offspring. Thus, genomic reprogramming factors are present in the cytoplasm of the GV oocyte and could facilitate cloning technology. This finding is also useful for research on the mechanisms involved in histone deacetylation and demethylation, even though histone methylation is thought to be epigenetically stable.
Bui
Hong-Thuy
HT
RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Kobe 650-0047, Japan. bthuy@konkuk.ac.kr
Wakayama
Sayaka
S
Kishigami
Satoshi
S
Kim
Jin-Hoi
JH
Van Thuan
Nguyen
N
Wakayama
Teruhiko
T
D000107:Acetylation; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D002457:Cell Extracts; D065150:Cellular Reprogramming; D033481:Chromosomes, Mammalian / Q000378:metabolism; D019976:Cloning, Organism; D003470:Culture Media; D003593:Cytoplasm / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006657:Histones / Q000378:metabolism; D034904:Lamin Type A / Q000235:genetics / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008745:Methylation; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*
D002457:Cell Extracts; D003470:Culture Media; D006657:Histones; D034904:Lamin Type A; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D008239:Lysine