title: Reprogramming of active and repressive histone modifications following nuclear transfer with rabbit mesenchymal stem cells and adult fibroblasts.
abstract: Following nuclear transfer (NT) the epigenetic state of a donor nucleus must be reprogrammed to an embryonic one. To evaluate the efficiency of nuclear reprogramming, we monitored the levels of histone H3 di/tri-methylated on lysine 4 (H3K4m2/3), a marker for transcriptionally active/permissive euchromatin, and of histone H3 tri-methylated on lysine 27 (H3K27m3), a modification associated with facultative heterochromatin, in embryos cloned using rabbit mesenchymal stem cells (MSC) and adult fibroblasts (RAF) isolated from the same animals. In vivo fertilized, in vitro cultured embryos served as controls. H3K27m3 was undetectable in all stages of control embryos except for weak staining in a few blastocyst cells. A similar situation was found in all NT embryos irrespective of the type of donor cells used, although both MSC and RAF stained substantially for H3K27m3. H3K4m2/3 levels were very high in one- and two-cell control embryos, but then decreased to reach a minimum at the eight-cell stage, and finally increased again to initial levels at the morula and blastocyst stage. Reprogramming of H3K4m2/3 differed remarkably among the different types NT embryos as well as between NT embryos and control embryos, and was apparently dependent on the type of donor cells. Interestingly, abnormal reprogramming of H3K4m2/3 was observed in NT embryos derived from both MSC and RAF, donor cell types with markedly different proliferation capacity. Our study demonstrates that the repressive chromatin modification, H3K27m3, is faithfully reprogrammed in NT embryos derived from MSC or RAF, while reprogramming of the activating chromatin modification, H3K4m2/3, is quite variable and does not reflect the situation observed in control embryos derived by fertilization.
authors.affiliation: Chair for Molecular Animal Breeding and Biotechnology, Department of Veterinary Sciences, LMU Munich , Oberschleissheim, Germany.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Brero
authors.forename: Ru
authors.initials: R
authors.lastname: Hao
authors.forename: Matthias
authors.initials: M
authors.lastname: Schieker
authors.forename: Matthias
authors.initials: M
authors.lastname: Wierer
authors.forename: Eckhard
authors.initials: E
authors.lastname: Wolf
authors.forename: Thomas
authors.initials: T
authors.lastname: Cremer
authors.forename: Valeri
authors.initials: V
authors.lastname: Zakhartchenko
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology; D049109:Cell Proliferation; D065150:Cellular Reprogramming; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D008745:Methylation; D053652:Nuclear Transfer Techniques; D011817:Rabbits
chemical_list: D006657:Histones