title: Comparison of two methods of in vivo gene transfer by electroporation.
abstract: 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.
authors.affiliation: 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
authors.forename: Hiroki
authors.initials: H
authors.lastname: Kubota
authors.forename: Yutaro
authors.initials: Y
authors.lastname: Hayashi
authors.forename: Yasue
authors.initials: Y
authors.lastname: Kubota
authors.forename: Kevin
authors.initials: K
authors.lastname: Coward
authors.forename: John
authors.initials: J
authors.lastname: Parrington
mesh_terms: 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
chemical_list: D001616:beta-Galactosidase