Production of germline chimeric chickens, with high transmission rate of donor‐derived gametes, produced by transfer of primordial germ cells

Abstract

Germline chimeric chickens were produced by transfer of primordial germ cells from White Leghorn to Barred Plymouth Rock, and vice versa. Blood was collected from stage 13-15 embryos and primordial germ cells were concentrated by Ficoll density gradient centrifugation. Approximately 200 primordial germ cells were injected into the bloodstream through the dorsal aorta of stage 14-15 recipient embryos from which blood had been drawn via the dorsal aorta prior to the injection. Intact embryos were also prepared as recipients for White Leghorns only. The manipulated embryos were cultured in recipient eggshells until hatching. Germline chimerism of the chickens reaching maturity was examined by mating them with Barred Plymouth Rocks and donor-derived offspring were identified based on their feather color. The efficiency of production of germline chimeras was 95% (19/20). When primordial germ cells were transferred from White Leghorn to Barred Plymouth Rock, the average frequency of donor-derived offspring was 81% for three male chimeras (96% for one female chimera), and it was approximately 3.5 times higher for transfer in the opposite direction (23% for 6 male chimeras). Removing blood from recipient embryos prior to primordial germ cell injection enhanced the frequency of donor-derived offspring by 10% in resulting male chimeras. Male chimeras produced donor-derived offspring more frequently (approximately 3.8 times) than female chimeras. Increases, decreases, or no changes were observed in the frequency of donor-derived offspring from the germline chimeras with increasing age.(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of mRNA for 3HADH in manipulated embryos to produce germline chimeric chickens

“…Chimeric chickens have previously been produced by the transfer of PGCs at the embryonic stage, and donor-derived offspring were obtained (Naito et al 1994;Furuta et al 2001).…”

“…It has been reported that exogenous genes could be introduced into the PGCs to produce a transgenic chicken (Naito et al 1998;Inada et al 1997;Eguma et al 1999;Fujihara 2000, van de Lavoir et al 2006). The transfer of PGCs from donor to recipient embryos is a commonly used technique to establish germline chimeric chickens (Naito et al 1994;Ono et al 1996;Yamaguchi et al 2000;Furuta et al 1999Furuta et al , 2001Furuta et al , 2007Furuta et al , 2008. The chickens produced by the transfer of PGCs were shown to be chimeric by using a progeny test (Naito et al 1994;Furuta et al 2001).…”

“…The transfer of PGCs from donor to recipient embryos is a commonly used technique to establish germline chimeric chickens (Naito et al 1994;Ono et al 1996;Yamaguchi et al 2000;Furuta et al 1999Furuta et al , 2001Furuta et al , 2007Furuta et al , 2008. The chickens produced by the transfer of PGCs were shown to be chimeric by using a progeny test (Naito et al 1994;Furuta et al 2001). The production of germlines of chimeric chicken is difficult because of the low hatchability of eggs with manipulated embryos.…”

Transfer of blood containing primordial germ cells between chicken eggs development of embryonic reproductive tract

“…Chimeric chickens have previously been produced by the transfer of PGCs at the embryonic stage, and donor-derived offspring were obtained (Naito et al 1994;Furuta et al 2001). Chimeric birds have also been produced by PGCs transfer from quail to chicken embryos (Ono et al 1996;.…”

“…PGCs circulate in the vascular system of the developing embryo and finally migrate into the germinal ridge (Fujimoto et al 1976;Kuwana 1993), where the cells differentiate into spermatogonia or oogonia. PGC transfer between embryonic blood vessels is a common technique for establishment of germ line chimeric chickens (Naito et al 1994;Ono et al 1996;Yamaguchi et al 2000;Furuta et al 2001), and it has been reported that exogenous genes may be introduced into PGCs to produce transgenic chicken (Naito et al 1998;Inada et al 1997;Eguma et al 1999;. For confirmation that transfer of PGCs is technically possible, quail PGCs have been H. Furuta (&) Á T. Sawada Á K. Nishikawa Á I. Yamamoto Á T. Yoshida Á M. Tanaka Department of Animal Science, Nippon Veterinary and Life Science University, Kyounan-cho 1-7-1, Musashino, Tokyo 180-8602, Japan e-mail: fhiroki@nvlu.ac.jp observed in chicken gonads using immunohistochemistry (Ono et al 1998), and the capability of producing chimeric chickens has been investigated by progeny testing (Naito et al 1994;Furuta et al 2001).…”