Comparison of two methods of in vivo gene transfer by electroporation.
OBJECTIVE
To evaluate two contrasting methods of in vivo gene transfer into testicular cells using electroporation, with regard to efficiency of transfer and damage to the testes.


METHODS
Controlled animal study.


METHODS
Research laboratory at a university medical school.


METHODS
8-10-week-old male mice.


METHODS
The reporter construct pCAGGS-LacZ consisting of a cytomegalovirus enhancer/chicken beta-actin promoter attached to the LacZ gene was introduced into the testes in vivo using electroporation. For eight weeks, the efficiency and extent of LacZ gene expression, and the extent to which the testis was damaged by the technique, were investigated.


METHODS
Beta-galactosidase activity resulting from expression of the LacZ transgene was verified by X-gal staining, and LacZ mRNA expression was determined by RT-PCR analysis. Potential disorders associated with seminiferous tubular sperm formation were evaluated using the Johnsen score.


RESULTS
Long-lasting beta-galactosidase activity was detected in spermatogenic cells up to eight weeks postelectroporation. Apparent damage to spermatogenesis was evident but was transient in nature and recovered with time; this plasticity was particularly evident following rete testes injection.


CONCLUSIONS
Injection into the rete testis appears to be more suitable for in vivo gene transfer by electroporation than direct intratesticular injection.
Kubota
Hiroki
H
Department of Surgical Medicine, Nephrourology, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601, Japan. kubochin@med.nagoya-cu.ac.jp
Hayashi
Yutaro
Y
Kubota
Yasue
Y
Coward
Kevin
K
Parrington
John
J
D000818:Animals; D017209:Apoptosis; D018274:Electroporation / Q000379:methods*; D018014:Gene Transfer Techniques; D007763:Lac Operon; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D013081:Sperm Motility; D013091:Spermatogenesis; D013737:Testis / Q000166:cytology / Q000502:physiology*; D001616:beta-Galactosidase / Q000235:genetics
D001616:beta-Galactosidase