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Testicular and ovarian gonocytes from 20-day incubated chicken embryos contribute to germline lineage after transfer into bloodstream of recipient embryos

¹ Transgenic Animal Research Center and ² Animal Genome Research Unit, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan and ³ Laboratory of Intellectual Fundamentals for Environmental Studies, National Institute for Environmental Studies, Tsukuba, Ibaraki 305-8506, Japan

Correspondence should be addressed to M Naito; Email: mnaito{at}affrc.go.jp

The present study was conducted to elucidate whether testicular and ovarian gonocytes obtained from 20-day incubated chicken embryos (stage 45) have the ability to migrate to the germinal ridges and contribute to germline lineage after transfer into the bloodstream of recipient embryos. Testicular and ovarian gonocytes were first identified as relatively large cells in a population of gonadal cells. The proportions of testicular and ovarian gonocytes in the total gonadal cells were 0.94 and 0.75% respectively, recognised as chicken vasa homologue-positive cells. Then, the dissociated gonadal cells obtained from 20-day incubated embryos containing testicular or ovarian gonocytes, with or without transfection, were transferred into recipient embryos. Expression of the introduced GFP gene was observed in the gonads of 6.5-day cultured recipient embryos (stage 30) in males and females, suggesting that the transferred testicular and ovarian gonocytes have the ability to migrate to the germinal ridges and enter the gonads. Furthermore, the presence of the donor-derived DNA was detected in the gonads of 20-day cultured recipient embryos in males and females, and also in the sperm samples obtained from the hatched male putative chimaeric chickens, suggesting that the transferred testicular and ovarian gonocytes were incorporated into the germline of chimaeric embryos and chickens. It is concluded that testicular and ovarian gonocytes obtained from 20-day incubated embryos have the ability to migrate to the germinal ridges after transfer into the bloodstream of recipient embryos, enter the gonads and contribute to the germline lineage of chimaeric embryos and chickens.