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Biology of Reproduction 67, 282-286 (2002)
© 2002 Society for the Study of Reproduction, Inc.

Regular Article

Quantitative Analysis of Gene Expression in Preimplantation Mouse Embryos Using Green Fluorescent Protein Reporter1

Serguei Yu. Medvedev2,a,b, Tomoyuki Tokunagaa, Richard M. Schultzc, Tsutomu Furukawaa, Takashi Nagaia, Manabu Yamaguchia, Misa Hosoea, Alexander F. Yakovlevb, Seiya Takahashia, and Yoshiaki Izaikea

a Department of Developmental Biology, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan b Department of Animal Biotechnology, All-Russian Institute of Animal Genetics and Breeding, Saint Petersburg 189620, Russia c Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6018

We have developed a method to monitor noninvasively, quantitatively, and in real-time transcription in living preimplantation mouse embryos by measuring expression of a short half-life form of enhanced green fluoresecent protein (EGFP) following microinjection of a plasmid-borne EGFP reporter gene. A standard curve was established by injecting known amounts of recombinant green fluorescent protein, and transcriptional activity was then determined by interpolating the amount of fluoresence in the DNA-injected embryos. This approach permitted multiple measurements in single embryos with no significant detrimental effect on embryonic development as long as light exposure was brief (<30 sec) and no more than two measurements were made each day. This method should facilitate analysis of the regulation of gene expression in preimplantation embryos; in particular, during the maternal-to-zygotic transition, and in other species in which limited numbers of embryos are available.

First decision: 20 December 2001.

1 This research was supported by the Japan Information Center of Science and Technology, the Organized Research Combination System Program, and by grant HD 22681 from the National Institutes of Health to R.M.S.

2 Correspondence: Serguei Yu. Medvedev, Department of Developmental Biology, National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan. FAX: 81 298 38 8635; sergyurimedvedev{at}netscape.net




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