title: Retrovirus-mediated in vitro gene transfer into chicken male germ line cells.
abstract: 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.
authors: BIOPHARM, Research Institute of Biopharmacy and Veterinary Drugs Ltd, 254 49 Jílové u Prahy, Czech Republic.
authors: Jirí
authors: J
authors: Kalina
authors: Filip
authors: F
authors: Senigl
authors: Alena
authors: A
authors: Micáková
authors: Jitka
authors: J
authors: Mucksová
authors: Jana
authors: J
authors: Blazková
authors: Haifeng
authors: H
authors: Yan
authors: Martin
authors: M
authors: Poplstein
authors: Jirí
authors: J
authors: Hejnar
authors: Pavel
authors: P
authors: Trefil
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D002645:Chickens; D005434:Flow Cytometry; D015870:Gene Expression; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D007248:Infertility, Male / Q000628:therapy / Q000662:veterinary; D008297:Male; D008856:Microscopy, Fluorescence; D013094:Spermatozoa / Q000502:physiology / Q000637:transplantation / Q000821:virology*; D013737:Testis / Q000166:cytology / Q000637:transplantation / Q000821:virology*; D014161:Transduction, Genetic / Q000379:methods*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics*
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins