Publisher : Asian Australasian Association of Animal Production Societies
DOI : 10.5713/ajas.2002.1227
Title & Authors
Migration Activity of Chicken Gonadal Primordial Germ Cells (gPGCs) and Post-transfer Localization of LacZ-transfected gPGCs in the Embryonic Gonads Jeong, D.K.; Han, J.Y.;
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 () 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 -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.
Cock Spermatozoa Serve as the Gene Vector for Generation of ransgenic Chicken (Gallus gallus),;;;;;;;
Asian-Australasian Journal of Animal Sciences, 2004. vol.17. 7, pp.885-891
Chang, I., A. Tajima, Y. Yasuda, T. Chikamune and T. Ohno. 1992. Simple method for isolation of primordial germ cells for chick embryos. Cell Biol. Int. Report 16:853-857.
Chang, I., D. K. Jeong, Y. H. Hong, T. S. Park, Y. K. Moon, T. Ohno and Y. J. Han. 1997. Production of germline chimeric chicken by transfer of cultured chick primordial germ cells. Cell Biol. Int. 21:495-499.
Hamburger, V. and H. L. Hamilton. 1951. A series of normal stages in the development of the chick embryo. J. Morph. 88:49-92.
Han, J. Y., R. N. Shoffner and K. S. Guise. 1994. Gene transfer by manipulation of primordial germ cell in the chicken. Asian-Aust. J. Anim. Sci. 7:427-434.
Han, J. Y., T. S. Park, Y. H. Hong, D. K. Jeong, J. N. Kim, K. D. Kim and J. M. Kim. 2002. Production of germline chimeras by transfer of chicken gonadal primordial germ cells maintained in vitro for an extended period. Theriogenology (in press)
Hong, Y. H., Y. H. Moon, D. K. Jeong and J. Y. Han. 1998. Improved transfection efficiency of chicken gonadal primordial germ cells for the production of transgenic poultry. Trans. Res. 7:247-252.
Love, J., C. Gribbin, C. Mather and H. Sang. 1994. Transgenic birds by DNA microinjection. Biotechnology 12:60-63.
Park, T. S. and J. Y. Han. 2000. Derivation and characterization of pluripotent embryonic germ cells in chicken. Mol. Reprod. Dev. 56:475-482.
Wentworth, B. C., H. Tsai, J. H. Lallett, D. S. Gonzales and G. Rajcic-Spasojevic. 1989. Manipulation of avian primordial germ cells and gonadal differentiation. Poultry Sci. 68:999-1010.