Positional changes of a pluripotency marker gene during structural reorganization of fibroblast nuclei in cloned early bovine embryos

Cloned bovine preimplantation embryos were generated by somatic cell nuclear transfer (SCNT) of bovine fetal fibroblasts with a silent copy of the pluripotency reporter gene GOF, integrated at a single site of a chromosome 13. GOF combines the regulatory Oct4/Pou5f1 sequence with the coding sequence for EGFP. EGFP expression served as a marker for pluripotency gene activation and was consistently detected in preimplantation embryos with 9 and more cells. Three-dimensional radial nuclear positions of GOF, its carrier chromosome territory and non-carrier homolog were measured in nuclei of fibroblasts, and of day 2 and day 4 embryos, carrying 2 to 9 and 15 to 22 cells, respectively. We tested, whether transcriptional activation was correlated with repositioning of GOF toward the nuclear interior either with a corresponding movement of its carrier chromosome territory 13 or via the formation of a giant chromatin loop. A significant shift of GOF away from the nuclear periphery was observed in day 2 embryos together with both carrier and non-carrier chromosome territories. At day 4, GOF, its carrier chromosome territory 13 and the non-carrier homolog had moved back toward the nuclear periphery. Similar movements of both chromosome territories ruled out a specific GOF effect. Pluripotency gene activation was preceded by a transient, radial shift of GOF toward the nuclear interior. The persistent co-localization of GOF with its carrier chromosome territory rules out the formation of a giant chromatin loop during GOF activation.

[1]  Volker J Schmid,et al.  Reprogramming of fibroblast nuclei in cloned bovine embryos involves major structural remodeling with both striking similarities and differences to nuclear phenotypes of in vitro fertilized embryos , 2014, Nucleus.

[2]  Thomas Cremer,et al.  Three-dimensional super-resolution microscopy of the inactive X chromosome territory reveals a collapse of its active nuclear compartment harboring distinct Xist RNA foci , 2014, Epigenetics & Chromatin.

[3]  E. Wolf,et al.  Fine mapping of genome activation in bovine embryos by RNA sequencing , 2014, Proceedings of the National Academy of Sciences.

[4]  K. Tanemura,et al.  Latrunculin A Treatment Prevents Abnormal Chromosome Segregation for Successful Development of Cloned Embryos , 2013, PloS one.

[5]  Lei Li,et al.  The maternal to zygotic transition in mammals. , 2013, Molecular aspects of medicine.

[6]  L. De Luca,et al.  Effects of oocyte quality, incubation time and maturation environment on the number of chromosomal abnormalities in IVF-derived early bovine embryos. , 2013, Reproduction, fertility, and development.

[7]  C. Boulesteix,et al.  3D-FISH analysis of embryonic nuclei in mouse highlights several abrupt changes of nuclear organization during preimplantation development , 2012, BMC Developmental Biology.

[8]  S. Bohlander,et al.  The radial nuclear positioning of genes correlates with features of megabase-sized chromatin domains , 2012, Chromosome Research.

[9]  J. Bridger.  Chromobility: the rapid movement of chromosomes in interphase nuclei. , 2011, Biochemical Society transactions.

[10]  Christopher H Eskiw,et al.  Farnesyltransferase inhibitor treatment restores chromosome territory positions and active chromosome dynamics in Hutchinson-Gilford progeria syndrome cells , 2011, Genome Biology.

[11]  Wael Tadros,et al.  The maternal-to-zygotic transition: a play in two acts , 2009, Development.

[12]  Rolf Ohlsson,et al.  Chromosome crosstalk in three dimensions , 2009, Nature.

[13]  L. Němcová,et al.  Gene expression during minor genome activation in preimplantation bovine development. , 2009, Theriogenology.

[14]  Thomas Cremer,et al.  Changes of higher order chromatin arrangements during major genome activation in bovine preimplantation embryos. , 2009, Experimental cell research.

[15]  E. Wolf,et al.  Reprogramming of active and repressive histone modifications following nuclear transfer with rabbit mesenchymal stem cells and adult fibroblasts. , 2009, Cloning and stem cells.

[16]  Thomas Cremer,et al.  Nuclear Architecture of Rod Photoreceptor Cells Adapts to Vision in Mammalian Evolution , 2009, Cell.

[17]  E. Wolf,et al.  Quantitative Monitoring of Pluripotency Gene Activation after Somatic Cloning in Cattle1 , 2007, Biology of reproduction.

[18]  Thomas Cremer,et al.  Epigenomic differentiation in mouse preimplantation nuclei of biparental, parthenote and cloned embryos , 2007, Chromosome Research.

[19]  Thomas Cremer,et al.  Radial chromatin positioning is shaped by local gene density, not by gene expression , 2007, Chromosoma.

[20]  Karl Rohr,et al.  Chromatin domains and the interchromatin compartment form structurally defined and functionally interacting nuclear networks , 2006, Chromosome Research.

[21]  J. S. Heslop-Harrison,et al.  Physical organization of the 1.709 satellite IV DNA family in Bovini and Tragelaphini tribes of the Bovidae: sequence and chromosomal evolution , 2006, Cytogenetic and Genome Research.

[22]  R. Eils,et al.  Three-Dimensional Maps of All Chromosomes in Human Male Fibroblast Nuclei and Prometaphase Rosettes , 2005, PLoS biology.

[23]  E. Wolf,et al.  Epigenetic reprogramming in mammalian nuclear transfer. , 2003, Differentiation; research in biological diversity.

[24]  J. Miyazaki,et al.  Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells , 2000, Nature Genetics.

[25]  H. Schöler,et al.  Germline‐specific expression of the Oct‐4/green fluorescent protein (GFP) transgene in mice , 1999, Development, growth & differentiation.

[26]  C. Popescu,et al.  Standard G-, Q-, and R-banded ideograms of the domestic sheep (Ovis aries): homology with cattle (Bos taurus). Report of the committee for the standardization of the sheep karyotype , 1999, Cytogenetic and Genome Research.

[27]  J. Williams,et al.  A high proportion of bovine blastocysts produced in vitro are mixoploid. , 1999, Biology of reproduction.

[28]  D. Hill,et al.  Isolation of chromosome-specific paints from high-resolution flow karyotypes of the sheep (Ovis aries) , 1997, Chromosome Research.

[29]  C. Epstein,et al.  Evidence for expression of the paternal genome in the two-cell mouse embryo , 1981, Nature.

[30]  Y Moreau,et al.  Microarray analysis reveals abnormal chromosomal complements in over 70% of 14 normally developing human embryos. , 2013, Human reproduction.

[31]  R. J. Mapletoft,et al.  Evaluation and classification of bovine embryos , 2013 .

[32]  T. Cremer,et al.  Nuclear architecture in developmental biology and cell specialisation. , 2011, Reproduction, fertility, and development.

[33]  E. Wolf,et al.  FISH on 3D preserved bovine and murine preimplantation embryos. , 2010, Methods in molecular biology.

[34]  T. Cremer,et al.  Chromosome territories. , 2010, Cold Spring Harbor perspectives in biology.

[35]  Thomas Cremer,et al.  Multicolor 3D fluorescence in situ hybridization for imaging interphase chromosomes. , 2008, Methods in molecular biology.

[36]  M. Singer.  Highly repeated sequences in mammalian genomes. , 1982, International review of cytology.