Transfection of oocytes and other types of ovarian cells in rabbits after direct injection into uterine arteries of adenoviruses and plasmid/liposomes.
Transfection of oocytes should be avoided in somatic gene therapy. However, several viral vectors including adenoviruses can transfect zona-pellucida-free eggs in vitro. During early stages of development, oocytes of postnatal ovaries lack the zona pellucida. Therefore, they may be susceptible to gene transfer and unintended toxic effects. The purpose of this study was to see whether the injection of adenoviruses (1 x 10(10) PFU) or plasmid (500 microg)/DOTMA:DOPE (1:2) liposomes directly into uterine arteries in pregnant rabbits leads to transfection of oocytes and other types of ovarian cells. LacZ and herpes simplex virus thymidine kinase (HSV-TK) were used as transgenes. It was found that both adenovirus and plasmid vectors transfected oocytes at the primordial and primary follicle stage when they were not protected by the zona pellucida, whereas no transfection was seen in oocytes surrounded by the zona pellucida. Efficient transfection of corpus luteum and granulosa cells was also detected by adenoviral and plasmid vectors. Transfection of oocytes and other ovarian cells was verified by X-gal staining and laser microdissection, followed by PCR analysis. HSV-TK gene transfer, followed by ganciclovir treatment, led to destruction of a significant number of oocytes, whereas HSV-TK gene transfer alone did not lead to toxic effects. It is concluded that the presence of a high concentration of adenovirus or plasmid vectors via the uterine artery may lead to transfection of zona-pellucida-free oocytes and other ovarian cells.
Laurema
A
A
A.I. Virtanen Institute, Kuopio, Finland.
Heikkilä
A
A
Keski-Nisula
L
L
Heikura
T
T
Lehtolainen
P
P
Manninen
H
H
Tuomisto
T T
TT
Heinonen
S
S
Ylä-Herttuala
S
S
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001158:Arteries; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D007269:Injections, Intra-Arterial; D008081:Liposomes; D009865:Oocytes / Q000378:metabolism*; D010053:Ovary / Q000166:cytology / Q000378:metabolism; D011817:Rabbits; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection / Q000379:methods*; D014599:Uterus / Q000098:blood supply; D001616:beta-Galactosidase / Q000235:genetics
D008081:Liposomes; D013937:Thymidine Kinase; D001616:beta-Galactosidase