title: Production of Interspecific Germline Chimeras via Embryo Replacement.
abstract: In avian species, primordial germ cells (PGCs) use the vascular system to reach their destination, the genital ridge. Because of this unique migratory route of avian germ cells, germline chimera production can be achieved via germ cell transfer into a blood vessel. This study was performed to establish an alternative germ cell-transfer system for producing germline chimeras by replacing an original host embryo with a donor embryo, while retaining the host extraembryonic tissue and yolk, before circulation. First, to test the migratory capacity of PGCs after embryo replacement, Korean Oge (KO) chick embryos were used to replace GFP transgenic chick embryos. Four days after replacement, GFP-positive cells were detected in the replaced KO embryonic gonads, and genomic DNA PCR analysis with the embryonic gonads demonstrated the presence of the GFP transgene. To produce an interspecific germline chimera, the original chick embryo proper was replaced with a quail embryo onto the chick yolk. To detect the gonadal PGCs in the 5.5-day-old embryonic gonads, immunohistochemistry was performed with monoclonal antibodies specific to either quail or chick PGCs, i.e., QCR1 and anti-stage-specific embryonic antigen-1 (SSEA-1), respectively. Both the QCR1-positive and SSEA-1-positive cells were detected in the gonads of replaced quail embryos. Forty percent of the PGC population in the quail embryos was occupied by chick extraembryonically derived PGCs. In conclusion, replacement of an embryo onto the host yolk before circulation can be applied to produce interspecies germline chimeras, and this germ cell-transfer technology is potentially applicable for reproduction of wild or endangered bird species.
authors.affiliation: Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Korea.
authors.forename: Hee Jung
authors.initials: HJ
authors.lastname: Choi
authors.affiliation: Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Korea.
authors.forename: Hyung Chul
authors.initials: HC
authors.lastname: Lee
authors.affiliation: Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Korea.
authors.forename: Kyung Soo
authors.initials: KS
authors.lastname: Kang
authors.affiliation: Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Korea.
authors.forename: Hyo Gun
authors.initials: HG
authors.lastname: Lee
authors.affiliation: Division of Animal Science, Faculty of Agriculture, Shinshu University, Nagano, Japan.
authors.forename: Tamao
authors.initials: T
authors.lastname: Ono
authors.affiliation: Laboratory for Early Embryogenesis, RIKEN Center for Developmental Biology, Kobe, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Nagai
authors.affiliation: Laboratory for Early Embryogenesis, RIKEN Center for Developmental Biology, Kobe, Japan.
authors.forename: Guojun
authors.initials: G
authors.lastname: Sheng
authors.affiliation: Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Korea Institute of Biomedical Science, Shinshu University, Nagano, Japan jaehan@snu.ac.kr.
authors.forename: Jae Yong
authors.initials: JY
authors.lastname: Han
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002642:Chick Embryo; D002678:Chimera / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004530:Egg Yolk / Q000502:physiology; D004622:Embryo, Mammalian; D005854:Germ Cells; D018095:Germ-Line Mutation / Q000235:genetics*; D006066:Gonads / Q000196:embryology; D049452:Green Fluorescent Proteins / Q000235:genetics; D007150:Immunohistochemistry; D016256:Lewis X Antigen / Q000235:genetics; D011784:Quail
chemical_list: D016256:Lewis X Antigen; D049452:Green Fluorescent Proteins; D004247:DNA
keywords: chicken; embryo replacement; interspecies germline chimera; primordial germ cell; quail