Transfection of oocytes and other types of ovarian cells in rabbits after direct injection into uterine arteries of adenoviruses and plasmid|[sol]|liposomes

doi:doi:10.1038/sj.gt.3301918

Gene Therapy (2003) 10, 580–584. doi:10.1038/sj.gt.3301918

Transfection of oocytes and other types of ovarian cells in rabbits after direct injection into uterine arteries of adenoviruses and plasmid/liposomes

A Laurema¹, A Heikkilä^1,2, L Keski-Nisula², T Heikura¹, P Lehtolainen¹, H Manninen³, T T Tuomisto¹, S Heinonen² and S Ylä-Herttuala^1,4,5

¹A.I. Virtanen Institute, Kuopio, Finland
²Department of Gynecology and Obstetrics, University of Kuopio, Kuopio, Finland
³Department of Radiology, University of Kuopio, Kuopio, Finland
⁴Department of Medicine, University of Kuopio, Kuopio, Finland
⁵Gene Therapy Unit, Kuopio University Hospital, Kuopio, Finland

Correspondence: S Ylä-Herttuala, Department of Molecular Medicine, A.I. Virtanen Institute, University of Kuopio, PO Box 1627, FIN-70211 Kuopio, Finland

Received 18 June 2002; Accepted 17 September 2002.

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Abstract

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 $times$ 10¹⁰ PFU) or plasmid (500 g)/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.