Oct4 distribution and level in mouse clones: consequences for pluripotency.
Somatic cell clones often fail at a developmental stage coincident with commencement of differentiation. The transcription factor Oct4 is expressed during cleavage stages and is essential for the differentiation of the blastocyst. Oct4 expression becomes restricted to the inner cell mass and epiblast. After gastrulation Oct4 is active only in germ cells and is silent in somatic cells. Here, Oct4 and an Oct4-GFP transgene were used as markers for which gene reprogramming could be directly related to the developmental potential of somatic cell clones. Cumulus cell clones initiated Oct4 expression at the correct stage but showed an incorrect spatial expression in the majority of blastocysts. The ability of clones to form outgrowths was reduced, and the outgrowths had low or even undetectable levels of Oct4 RNA or GFP. The quality of GFP signals in blastocysts correlated with the ability to generate outgrowths that maintain GFP expression and the frequency of embryonic stem (ES) cell derivation. Abnormal Oct4 expression in clones is either directly or indirectly caused by reprogramming errors and is indicative of a general failure to reset the genetic program. The abnormal Oct4 expression may be associated with aberrant expression of other crucial developmental genes, leading to abnormalities at various embryonic stages. Regardless of other genes, the variations observed in Oct4 levels alone account for the majority of failures currently observed for somatic cell cloning.
Boiani
Michele
M
Germline Development Group, Center for Animal Transgenesis and Germ Cell Research, The School of Veterinary Medicine, University of Pennsylvania, New Bolton Center, Kennett Square, Pennsylvania 19348, USA.
Eckardt
Sigrid
S
Schöler
Hans R
HR
McLaughlin
K John
KJ
D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D002454:Cell Differentiation; D002455:Cell Division; D002467:Cell Nucleus / Q000502:physiology; D002470:Cell Survival; D002999:Clone Cells; D004268:DNA-Binding Proteins / Q000502:physiology*; D004622:Embryo, Mammalian; D005260:Female; D005307:Fertilization in Vitro; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D050814:Octamer Transcription Factor-3; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000502:physiology
D004268:DNA-Binding Proteins; D008164:Luminescent Proteins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D012333:RNA, Messenger; D014157:Transcription Factors; D049452:Green Fluorescent Proteins