Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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BOR - Papers in Press, published online ahead of print June 30, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.030908
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Submitted April 15, 2004
Returned for revision May 7, 2004
Accepted June 9, 2004

Reproductive Technology


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

Katsutoshi Niwa , Riya Takano , Yayoi Obata , Hitoshi Hiura , Junichi Komiyama , Hidehiko Ogawa , and Tomohiro Kono *

* To whom correspondence should be addressed. E-mail: tomohiro{at}nodai.ac.jp.

Abstract
Mouse parthenotes result in embryonic death before 10 days of gestation, but parthenogenetic embryos (ng/fg PE) that contain haploid sets of genomes from non-growing (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.

Key words: Embryo • Gamete Biology • Developmental biology • Gene regulation • Oocyte development


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