The growth arrest genes gas5, gas6, and CHOP-10 (gadd153) are expressed in the mouse preimplantation embryo.
The gas and gadd family of genes, known collectively as the growth arrest genes, are associated with the negative control of mammalian cell growth. The steady-state levels of their mRNAs are increased by three to fivefold when exponentially multiplying cells are exposed to a variety of stresses including inadequate nutrition or the removal of serum. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to analyze growth arrest gene expression in the preimplantation mouse embryo. The gas5, gas6, and CHOP-10 (gadd153, Ddit3) genes were expressed from the eight-cell stage onward. The gas2 and gas3 genes associated with apoptosis were not expressed. Embryos were cultured in kSOM medium and a semiquantitative RT-PCR method was used to measure the relative gene expression using beta-actin mRNA as a reference. The ratio of gas5 to beta-actin mRNA was high at the eight-cell stage and fell three to fivefold during development. The decline in the gas5:beta-actin ratio corresponded to the activation of true cell growth (cytokinesis). The gas6:beta-actin ratio was low at the eight-cell stage and increased by twofold as the blastocyst formed. CHOP-10 was expressed at a constant level throughout development. Embryos that had developed in vivo were compared with the equivalent blastocyst-stage embryos cultured in kSOM medium. There were no significant differences in the ratio of CHOP-10, gas5, or gas6 mRNAs relative to beta-actin. These results suggest that these genes are expressed as part of normal early embryonic development. The potential roles of the growth arrest genes are discussed.
Fleming
J V
JV
Rowett Research Institute, Aberdeen, Scotland, United Kingdom.
Fontanier
N
N
Harries
D N
DN
Rees
W D
WD
D016475:3T3 Cells; D000199:Actins / Q000096:biosynthesis; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D022762:CCAAT-Enhancer-Binding Proteins; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D036341:Intercellular Signaling Peptides and Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D012313:RNA / Q000032:analysis; D051742:Transcription Factor CHOP; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic; D000099255:Growth Arrest-Specific Protein 6
D000199:Actins; D022762:CCAAT-Enhancer-Binding Proteins; D004268:DNA-Binding Proteins; D036341:Intercellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D011506:Proteins; D012313:RNA; D051742:Transcription Factor CHOP; D014157:Transcription Factors; D000099255:Growth Arrest-Specific Protein 6; C497250:Ddit3 protein, mouse