Transcriptomics,Genomics

Dataset Information

39

First round of DNA replication is essential for transcriptional reprogramming of somatic nuclei in mouse oocytes


ABSTRACT: Many of the structural and mechanistic requirements of oocyte-mediated nuclear reprogramming remain elusive. Previous accounts that transcriptional reprogramming of somatic nuclei in mouse zygotes may be complete in 24-36 hours, far more rapidly than in other reprogramming systems, raise the question of whether the mere exposure to the activated mouse ooplasm is sufficient to enact reprogramming in a nucleus. We therefore prevented DNA replication and cytokinesis, which ensue after nuclear transfer, in order to assess their requirement for transcriptional reprogramming of the key pluripotency genes Oct4 (Pou5f1) and Nanog in cloned mouse embryos. Using transcriptome and allele-specific analysis, we observed that hundreds of mRNAs, but not Oct4 and Nanog, became elevated in nucleus-transplanted oocytes without DNA replication. Progression through the first round of DNA replication was essential but not sufficient for transcriptional reprogramming of Oct4 and Nanog, whereas cytokinesis and thereby cell-cell interactions were dispensable for transcriptional reprogramming. Responses similar to clones also were observed in embryos produced by fertilization in vitro. Our results link the occurrence of reprogramming to a previously unappreciated requirement of oocyte-mediated nuclear reprogramming, namely DNA replication. Nuclear transfer alone affords no immediate transition from a somatic to a pluripotent gene expression pattern unless DNA replication is also in place. This study is therefore a resource to appreciate that the quest for always faster reprogramming methods may collide with a limit that is dictated by the cell cycle. Overall design: 13 samples were analyzed. Aph-IVF: Aphidicolin not treated, in vitro fertilization, 1 biological rep Aph-IVF96: Aphidicolin not treated, in vitro fertilization (96h), 1 biological rep Aph-IVFBla: Aphidicolin not treated, in vitro fertilization blastocyst, 1 biological rep Aph+IVF: Aphidicolin treated, in vitro fertilization, 1 biological rep Aph+IVF96: Aphidicolin treated, in vitro fertilization (96h), 1 biological rep Aph+IVFBla: Aphidicolin treated, in vitro fertilization blastocyst, 1 biological rep Aph-SCNT: Aphidicolin not treated, somatic cell nuclear transfer, 1 biological rep Aph-SCNT96: Aphidicolin not treated, somatic cell nuclear transfer (96h), 1 biological rep Aph-SCNTBla: Aphidicolin not treated, somatic cell nuclear transfer blastocyst, 1 biological rep Aph+SCNTBla: Aphidicolin treated, somatic cell nuclear transfer blastocyst, 1 biological rep B6C3F1-Ooc: Aphidicolin not treated, B6C3F1 oocytes, 1 biological rep B6C3F1-Cum: Aphidicolin not treated, B6C3F1 cumulus cells, 2 biological rep

INSTRUMENT(S): Illumina MouseRef-8 v2.0 expression beadchip

SUBMITTER: Marcos J Araúzo-Bravo  

PROVIDER: GSE53497 | GEO | 2016-12-17

SECONDARY ACCESSION(S): PRJNA232136

REPOSITORIES: GEO

Similar Datasets

| GSE110851 | GEO
2011-08-16 | GSE23654 | GEO
2011-08-15 | E-GEOD-23654 | ArrayExpress
2013-07-11 | E-GEOD-36468 | ArrayExpress
| GSE59073 | GEO
| GSE89279 | GEO
2012-08-10 | E-GEOD-38711 | ArrayExpress
| GSE94267 | GEO
| GSE95311 | GEO
| PRJNA387179 | ENA