This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kalina, J. Articles by Trefil, P. Search for Related Content PubMed PubMed Citation Articles by Kalina, J. Articles by Trefil, P.

Retrovirus-mediated in vitro gene transfer into chicken male germ line cells

Jií Kalina, Filip enigl¹, Alena Miáková, Jitka Mucksová, Jana Blaková¹, Haifeng Yan², Martin Popltein, Jií Hejnar¹ and Pavel Trefil

BIOPHARM, Research Institute of Biopharmacy and Veterinary Drugs Ltd, 254 49 Jílové u Prahy, Czech Republic, ¹ Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Vídeská 1083, CZ-14220 Prague 4, Czech Republic and ² HIAVS, Hunan Institute of Animal and Veterinary Science, Quantang, Changsha 410131, Hunan, China

Correspondence should be addressed to J Hejnar; Email: P Trefil; Email: trefil{at}bri.cz

Chicken testicular cells, including spermatogonia, transplanted into the testes of recipient cockerels sterilized by repeated -irradiation repopulate the seminiferous epithelium and resume the exogenous spermatogenesis. This procedure could be used to introduce genetic modifications into the male germ line and generate transgenic chickens. In this study, we present a successful retroviral infection of chicken testicular cells and consequent transduction of the retroviral vector into the sperm of recipient cockerels. A vesicular stomatitis virus glycoprotein G-pseudotyped recombinant retroviral vector, carrying the enhanced green fluorescent protein reporter gene was applied to the short-term culture of dispersed testicular cells. The efficiency of infection and the viability of infected cells were analyzed by flow cytometry. No significant CpG methylation was detected in the infected testicular cells, suggesting that epigenetic silencing events do not play a role at this stage of germ line development. After transplantation into sterilized recipient cockerels, these retrovirus-infected testicular cells restored exogenous spermatogenesis within 9 weeks with approximately the same efficiency as non-infected cells. Transduction of the reporter gene encoding the green fluorescent protein was detected in the sperms of recipient cockerels with restored spermatogenesis. Our data demonstrate that, similarly as in mouse and rat, the transplantation of retrovirus-infected spermatogonia provides an efficient system to introduce genes into the chicken male germ line.

This article has been cited by other articles:

				J. G. Jung, Y. M. Lee, J. N. Kim, T. M. Kim, J. H. Shin, T. H. Kim, J. M. Lim, and J. Y. Han The reversible developmental unipotency of germ cells in chicken Reproduction, January 1, 2010; 139(1): 113 - 119. [Abstract] [Full Text] [PDF]