Expression of a fluorescent recombinant form of sperm protein phospholipase C zeta in mouse epididymal sperm by in vivo gene transfer into the testis

To use in vivo gene transfer into the testis by electroporation to express a fluorescent recombinant form of a testis-specific gene in the mature epididymal sperm of mice and thus study the pattern of gene localization. Controlled animal study. Research laboratory at the University of Oxford. Four-...

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Bibliographic Details
Published in:	Fertility and sterility Vol. 85 Suppl 1; p. 1281
Main Authors:	Coward, Kevin, Kubota, Hiroki, Hibbitt, Olivia, McIlhinney, Jeff, Kohri, Kenjiro, Parrington, John
Format:	Journal Article
Language:	English
Published:	United States 01-04-2006
Subjects:	Animals Bacterial Proteins - genetics Bacterial Proteins - metabolism Cells, Cultured Electroporation - methods Epididymal Secretory Proteins - biosynthesis Epididymal Secretory Proteins - genetics Gene Expression Regulation - physiology Gene Transfer Techniques Luminescent Proteins - genetics Luminescent Proteins - metabolism Male Mice Mice, Transgenic Microscopy, Fluorescence Phosphoinositide Phospholipase C Recombinant Fusion Proteins - metabolism Spermatozoa - metabolism Testis - metabolism Tissue Distribution Type C Phospholipases - genetics Type C Phospholipases - metabolism
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Summary:	To use in vivo gene transfer into the testis by electroporation to express a fluorescent recombinant form of a testis-specific gene in the mature epididymal sperm of mice and thus study the pattern of gene localization. Controlled animal study. Research laboratory at the University of Oxford. Four- to 6-week-old male mice. Phospholipase C zeta (PLCzeta), the putative mammalian egg activation factor, was fused to enhanced yellow fluorescent protein (EYFP), and in vivo gene transfer by electroporation was used to introduce this transgene (PLCzeta-EYFP) into mouse testis. Transgene expression in testis and sperm were analyzed at 20 and 40 days after electroporation. Transgene expression in testis and epididymal sperm was analyzed by fluorescence microscopy and an excitation light source suitable for EYFP. Phospholipase C zeta-EYFP was successfully expressed in epididymal sperm when analyzed 40 days after gene transfer and was localized to the head and midpiece regions. Our results provide the first demonstration that in vivo gene transfer can be used to study the localization of proteins in mature sperm and that this represents a powerful new technique for studying male infertility and gene function in sperm.
ISSN:	1556-5653
DOI:	10.1016/j.fertnstert.2005.12.012