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Nuclei of Oocytes Derived from Mouse Parthenogenetic Embryos Are Competent to Support Development to Term¹

Department of BioScience,³ Tokyo University of Agriculture, Tokyo 156-8502, Japan Bio-oriented Technology Research Advancement Institution (BRAIN),⁴ Tokyo 105-0001, Japan

Mouse parthenotes result in embryonic death before 10 days of gestation, but parthenogenetic embryos (ng/fg PE) that contain haploid sets of genomes from nongrowing (ng) oocytes derived from newborn fetuses and fully grown (fg) oocytes derived from adults can develop into 13.5-day-old fetuses. This prolonged development is due to a lack of genomic imprinting in ng oocytes. Here, we show maternal genomes of oocytes derived from ng/fg PE are competent to support normal development. After 28 days of culture, the ovaries from ng/fg PE grew as well as the controls, forming vesicular follicles with follicular antrums. The oocytes collected from the developed follicles were the same size as those of the controls. To determine whether maternal primary imprinting had been established in the oocytes derived from ng/fg PE, we examined the DNA methylation status in differentially methylated regions of three imprinted genes, Igf2r, Lit1, and H19. The results showed that maternal-specific modifications were imposed in the oocytes derived from ng/fg PE. Further, to assess nuclear competence to support development, we constructed matured oocytes containing a haploid genome derived from ng/fg PE oocytes by serial nuclear transfer. After in vitro fertilization and culture and embryo transplantation into recipients, two live pups were obtained. One developed normally to a fertile adult. These results revealed that oocytes derived from ng/fg PE can be normally imprinted during oogenesis and acquire competence to participate in development as female genomes.

¹ Supported by grants from the Bio-Oriented Technology Research Advancement Institution (BRAIN), Japan, and the Ministry of Education, Science, Culture and Sports of Japan.

² Correspondence: Tomohiro Kono, Department of BioScience, Tokyo University of Agriculture, 1-1-1, Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan. FAX: 81 3 5477 2543; tomohiro{at}nodai.ac.jp

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