[Effects of the gene transfer into sperm mediated by liposomes on sperm motility and fertilization in vitro].
To clarify the problems of gene transfer in the fertility clinic, we investigated the safety and effectiveness of the gene transfer of mouse sperm using the lipofection method. We used the pCAGGS-lacZ plasmid connected with cytomegarovirus enhancer/chicken beta-actin promoter as an adventitious gene. After incubation of the DNA-liposome complex and mouse sperm of epididymis in the medium, we investigated the sperm motility after gene transfer, in vitro fertilization rate of the gene transferred sperm, and gene expression of the fertilized ovum and fetus. Sperm motility was lowered significantly with the increase in DNA density. The fertilization rate for in vitro fertilization was lowered significantly with the increase in DNA density. Gene expression was observed in some of the fertilized ova. In the fetus 12.5 days after embryo transfer, gene transfer was confirmed by the PCR method, but we could not find gene expression by X-gal staining. We proved that the sperm with a gene transferred by the lipofection method could fertilize the ovum and transfer adventitious gene to ova. We speculate that sperm motility and fertility rate were lowered because the introduction of DNA and the DNA-liposome complex destroyed parts of the cell. Development of a sperm fractionation method for DNA transfer and establishment of a method for effective and safe DNA transfer are awaited.
Sasaki
S
S
Department of Urology, Nagoya City University Medical School.
Kojima
Y
Y
Kubota
H
H
Tatsura
H
H
Hayashi
Y
Y
Kohri
K
K
D000818:Animals; D005260:Female; D005307:Fertilization in Vitro; D018014:Gene Transfer Techniques; D008081:Liposomes; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008815:Mice, Inbred Strains; D013081:Sperm Motility / Q000502:physiology*; D013094:Spermatozoa
D008081:Liposomes