In vitro manipulation of early mouse embryos induces HIV1-LTRlacZ transgene expression.
We report here that the transcriptional activity of early mouse embryos is affected by their manipulation and culture in vitro, using transgenic embryos that express the reporter gene lacZ. We examined the pattern of expression of the lacZ gene fused to the human immunodeficiency virus type 1 long terminal repeat during the preimplantation stages. Transgene expression is induced as early as the two-cell stage in embryos developed in vitro, while there is no constitutive expression at the same stage in embryos developed in vivo. We have established a relation between this inducible expression occurring in vitro and an oxidative stress phenomenon. Indeed, when the culture medium is supplemented with antioxidants such N-acetyl-cysteine or CuZn-superoxide dismutase the transgene expression is markedly reduced. We also present evidence that the transgene expression in vitro coincides with the onset of the embryonic genome activation as attested by the synthesis of the 70 x 10(3) M(r) protein complex. Therefore, this transgene expression could prove to be a useful tool in our understanding of the molecular mechanisms involved in this crucial developmental event.
Vernet
M
M
Institut Cochin de Génétique Moléculaire, Laboratoire de Génétique et Pathologie Expérimentales-CJF INSERM 90-03, Paris, France.
Cavard
C
C
Zider
A
A
Fergelot
P
P
Grimber
G
G
Briand
P
P
D000818:Animals; D001345:Autoradiography; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation / Q000502:physiology*; D005814:Genes, Viral / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics*; D007763:Lac Operon / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic / Q000196:embryology / Q000235:genetics; D010084:Oxidation-Reduction; D014158:Transcription, Genetic