Deficiency in recapitulation of stage-specific embryonic gene transcription in two-cell stage cloned mouse embryos.
One possible explanation to account for the ability to clone animals by somatic cell nuclear transfer is that the donor genome is reprogrammed by the oocyte to recapitulate a normal embryonic pattern of gene expression. Mouse embryos display transient transcriptional induction of a group of genes (TIGs) at the mid two-cell stage, the first major transcriptional output of the embryonic genome, uniquely suited for evaluating whether the oocyte directs correct and efficient recapitulation of an embryo stage-specific gene expression program before any in vitro selection occurs. We analyzed the expression of eight TIGs in two-cell stage clones prepared with cumulus cell nuclei. One failed to be transcribed, and seven others were transcribed, but supported significantly reduced mRNA expression. The reduction ranged from 1.6- to 17-fold at the mid two-cell stage, and 1.5- to 13-fold for the late two-cell stage. Five genes were not expressed in the donor cells, and these displayed the most pronounced deficiencies in expression. Two genes were expressed in cumulus cell donors, and supported progressive accumulation of their mRNAs in clones during this period, albeit at reduced rates. One other gene expressed in cumulus cells was not activated in clones. Although a significant proportion of these genes is reactivated in two-cell stage clones, this recapitulation is grossly imperfect, occurs at a substantially reduced level, and even fails entirely to occur for some genes. Thus, the oocyte is incapable of efficiently directing the recapitulation of early embryonic stage-specific gene expression.
Vassena
Rita
R
The Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, USA.
Han
Zhiming
Z
Gao
Shaorong
S
Latham
Keith E
KE
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D019976:Cloning, Organism; D004622:Embryo, Mammalian / Q000378:metabolism*; D047108:Embryonic Development / Q000235:genetics*; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D014158:Transcription, Genetic
D012333:RNA, Messenger