Retrovirus-mediated in vitro gene transfer into chicken male germ line cells
Language English Country England, Great Britain Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
17709563
DOI
10.1530/rep-06-0233
PII: 134/3/445
Knihovny.cz E-resources
- MeSH
- Gene Expression MeSH
- Microscopy, Fluorescence MeSH
- Genetic Vectors administration & dosage genetics MeSH
- Animals, Genetically Modified MeSH
- Cells, Cultured MeSH
- Chickens * MeSH
- Infertility, Male therapy veterinary MeSH
- Flow Cytometry MeSH
- Spermatozoa physiology transplantation virology MeSH
- Testis cytology transplantation virology MeSH
- Transduction, Genetic methods MeSH
- Vesicular stomatitis Indiana virus genetics MeSH
- Green Fluorescent Proteins genetics MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- enhanced green fluorescent protein MeSH Browser
- Green Fluorescent Proteins MeSH
Chicken testicular cells, including spermatogonia, transplanted into the testes of recipient cockerels sterilized by repeated gamma-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.
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