Nuclear profiles of H3 histones trimethylated on Lys27 in bovine (Bos taurus) embryos obtained after in vitro fertilization or somatic cell nuclear transfer.
Histone H3 trimethylation on lysine 27 is one of the histone modifications associated with chromatin of silenced regions. H3K27me3 labeling is initially asymmetrical between pronuclei in mammalian embryos, and then it is remodeled during early development. However, in mouse embryos obtained after somatic cell nuclear transfer (SCNT), H3K27me3 histones inherited from the somatic female cell and associated with X chromosome inactivation have been reported to escape remodeling. Using immunostaining, we investigated the remodeling of H3K27me3 in Bos taurus embryos obtained after in vitro fertilization (IVF) and SCNT. In this species, transfer-induced chromatin remodeling can be clearly separated from embryonic genome activation (EGA), which occurs at the 8-16-cell stage, and cloning by SCNT is 10 times more successful than in the mouse. In early IVF bovine embryos, dense H3K27me3 labeling was localized in the pericentric heterochromatin as recently described in the mouse. Labeling was however unevenly distributed up to the 8-cell stage, suggesting that the parental genomes partitioned before EGA. In female IVF blastocysts, a somatic-like female profile appeared in 21% of the trophoblast cells. This profile, which had one major nuclear H3K27me3 patch, the putative inactive X chromosome (Xi), was absent in male blastocysts. In contrast, the somatic-like female H3K27me3 profile was observed in the majority of the nuclei of female bovine SCNT embryos before EGA. At the 8-16-cell stage, this profile was transiently replaced by pericentric-like labeling in most nuclei. Immunostaining of mitotic chromosomes suggested that the ratio of H3K27me3 labeling in pericentric heterochromatin vs. euchromatin was then rapidly altered. Finally, Xi-like H3K27me3 staining appeared again in trophoblast cells in female SCNT blastocysts. These results suggest a role for EGA in H3K27me3 remodeling, which affects the heterochromatin inherited from the donor cell or produced during development.
Breton
Amandine
A
INRA, ENVA UMR 1198 Biologie du Développement et Reproduction, France.
LE Bourhis
Daniel
D
Audouard
Christophe
C
Vignon
Xavier
X
Lelièvre
Jean-Marc
JM
D000818:Animals; D002417:Cattle / Q000196:embryology* / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D042002:Chromatin Assembly and Disassembly / Q000502:physiology; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D006570:Heterochromatin / Q000378:metabolism; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D006822:Hybrid Cells / Q000378:metabolism / Q000502:physiology; D008239:Lysine / Q000378:metabolism; D008297:Male; D055442:Metabolome; D008745:Methylation; D051379:Mice; D053652:Nuclear Transfer Techniques; D011247:Pregnancy
D006570:Heterochromatin; D006657:Histones; D008239:Lysine