In vivo gene transfer to mouse spermatogenic cells using green fluorescent protein as a marker

The Journal of Experimental Zoology
Y YamazakiK Imoto

Abstract

Combination of the DNA injection into seminiferous tubules and the subsequent in vivo electroporation (EP) has become an efficient and convenient assay system for spermatogenic-specific gene expression during spermatogenesis of mice. In this study, we made methodological modifications to enhance the transfection efficiency, and evaluated the possibility of this technique to generate transgenic offspring using green fluorescent protein (GFP) as a marker. After the in vivo gene transfer, GFP expression could be monitored easily and repeatedly on the surface of the testis of live mice under fluorescent microscopy. The serial sections of the transfected testis revealed that transient expression of GFP was extended even in the innermost region of the testis uniformly, but confined to spermatogenic cells and Sertoli cells within the seminiferous tubules. Furthermore, long-lasting GFP expression could be detected in the spermatogenic cells even 2 months after EP. Natural mating with normal adult females revealed that 65% of the transfected males maintained fertilizable ability and could generate their offspring normally. Germ-line transmission of the GFP vector to the offspring was checked under fluorescent microscopy, but no transgen...Continue Reading

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Related Concepts

Gene Transfer Techniques
Reproduction
Testis
Green Fluorescent Proteins
Spermiogenesis
Founder Mice, Transgenic
Y-Chromosome-Bearing Sperm
Luminescent Proteins
Metazoa
Mouse, Swiss

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