In vivo transfection of the mouse vas deferens.
To explore the possibility that specific characteristics of the epithelium of the male tract can be modified, transfections of the mouse vas deferens have been performed using in vivo injections of cationic DNA/liposome complexes. Gene transfer was done employing the reporter genes pEGFP-C1 encoding Green Fluorescent Protein (GFP) and pCMV-nls-beta encoding the nuclear beta-Galactosidase (beta-Gal). Foreign gene expression reached a maximum of 6.8% in the epithelial cells of the vas after treatment with the nuclear beta-Gal gene construction and of 13.3% after employing the GFP gene construction. Expression of the GFP gene appeared from one week up to three months following injection, and it appeared as patches of modified cells along the epithelium. Results from immunocytochemistry and Western Blotting support the conclusion that transfection of epithelial cells was achieved. We have also transfected the vas using gene constructions that express secretory proteins--specifically, the reporter system pSEAP-control that expresses a secretory form of human placental alkaline phosphatase, and the pGFP-Ctk-37 that expresses a secretion form of GFP. In both cases, the fluids expressed from the transfected vas showed a significant increase of alkaline phosphatase activity after pSEAP transfection and the presence of GFP protein when pGFP-Ctk-37 gene construction was employed. Our results indicate that the vas can be transfected in vivo using liposomes as vectors of foreign genes and that the vas fluid contents can be modified.
Valenzuela
Mario
M
Centro de Investigaciones Biológicas, CSIC, Velázquez 144, 28006 Madrid, Spain.
Relloso
Miguel
M
Esponda
Pedro
P
D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival; D004247:DNA / Q000008:administration & dosage; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter / Q000235:genetics*; D049452:Green Fluorescent Proteins; D006801:Humans; D007150:Immunohistochemistry; D008081:Liposomes / Q000008:administration & dosage; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D008297:Male; D051379:Mice; D014162:Transfection / Q000379:methods*; D014649:Vas Deferens / Q000166:cytology / Q000378:metabolism*; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D008081:Liposomes; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase