Real time imaging of transcriptional activity in live mouse preimplantation embryos using a secreted luciferase.
Study of gene expression kinetics during preimplantation mammalian development is difficult because of the limited amount of material and the usually destructive, static nature of molecular analyses. We describe continuous, noninvasive monitoring of gene expression in preimplantation embryos by using a secreted luminescent reporter, Vargula luciferase. Transgene expression profiles were followed by assaying aliquots of culture medium or by direct visualization of Vargula luciferase secretion from living embryos in real time through photon imaging. With this approach, it is possible to observe epigenetic modulations of gene expression and to link this over time to the developmental capacity of individual embryos. In addition, by developing a strategy where expression from integrated transgenes is enhanced relative to that from nonintegrated DNA, we provide evidence that rapid detection of transgene integration prior to the blastocyst stage should be possible. Thus, imaging of Vargula luciferase secretion may also be useful in the early screening of embryos, for example, in the production of transgenic livestock.
Thompson
E M
EM
Unité de Biologie du Développement, Institut National de la Recherche Agronomique, Jouy-en Josas, France.
Adenot
P
P
Tsuji
F I
FI
Renard
J P
JP
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002970:Cleavage Stage, Ovum / Q000502:physiology; D047108:Embryonic Development; D005260:Female; D018507:Gene Expression Regulation, Developmental; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D006358:Hot Temperature; D006877:Hydroxamic Acids / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008845:Microinjections; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D013053:Spectrophotometry; D014158:Transcription, Genetic
D018840:HSP70 Heat-Shock Proteins; D006877:Hydroxamic Acids; D012333:RNA, Messenger; C012589:trichostatin A; D008156:Luciferases