Abstract
A powerful tool for chicken transgenesis could be established by employing a germline chimera production through primordial germ cell transplantation. This study was conducted to examine whether foreign gene-transfected gonadal primordial germ cells (gPGCs) have a migration activity into the gonad after transfer to recipient embryos. In Experiment 1, gPGCs of Korean Ogol Chicken were retrieved from 5.5-day-old embryos and subsequently transferred to the dorsal aorta of 2.5-day-old White Leghorn embryos after being labeled with PKH26 fluorescent dye. To confirm migration activity after transplantation, recipient embryos were sacrificed and examined on 3 days after transfer. Sex determination was concomitantly undertaken to examine whether sex of recipient embryos could affect the migration activity of gPGCs. All of embryonic gonads examined showed positive signals with PKH26 fluorescence and W-chromosome specific band by polymerase chain reaction (PCR) was detected in male embryos when gPGCs with ZW chromosome were transferred to recipient embryos. In Experiment 2, retrieved gPGCs were transfected with LacZ gene-containing cytomegalovirus promoter (pCMVβ) by electroporation and subsequently transferred to recipient embryos. LacZ gene expression was identified in the gonads of 6 or 10-day-old recipient embryos and hatched-chicks. A total of 20 embryos and 12 hatched-chicks were examined and 11 of them (10 embryos and one hatched chicken; 11/32=34.4%) expressed β-galactosidase, a marker substance of LacZ gene. The results of this study demonstrated that foreign gene-transfected gPGCs can migrate and settle down into the gonad after being transferred into the blood vessel of the recipient embryos. This established technique will contribute to developing a peer biotechnology for transgenic chicken. (Asian-Aust. J. Anim. Sci. 2002. Vol 15, No. 9 : 1227-1231)
Highlights
Production of transgenic chicken has an unlimited value in various fields of animal biotechnology
The results of this study demonstrated that gonadal primordial germ cells (gPGCs) transfected with pCMVβ by electroporation can migrate and settle down into the gonads after being transferred into the blood vessel of the recipient embryos
Localization and expression of LacZ gene and its maker substance, βgalactosidase, were confirmed in the gonads retrieved from recipient embryos or hatched chicks developed from recipient embryos
Summary
Production of transgenic chicken has an unlimited value in various fields of animal biotechnology. The use of primordial germ cells (PGCs) is the most powerful tool for improving the efficacy of transgenic technology and PGCs of several types, which were retrieved from the germinal crescent, blood or gonad in embryos of different stages, have been used to date. Collection of PGC from embryonic gonad yielded a number of critical benefits for the production of germline chimera. Primordial germ cells can be and efficiently collected from the embryonic gonads, compared with those from the germinal crescent or embryonic blood. It was recently reported that gonadal primordial germ cells (gPGCs) cultured in vitro have an activity of embryonic germ cells (Park and Han, 2000) and that the transfer of cultured gPGCs into embryos can induce germline chimerism (Chang et al, 1997). All of previous data strongly demonstrated the feasibility of gPGCs for improving chicken transgenic technology
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