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Mol. Hum. Reprod. Advance Access published online on March 30, 2009
Molecular Human Reproduction, doi:10.1093/molehr/gap026
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© The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

In-vivo gene transfer induces transgene expression in cells and secretions of the mouse cauda epididymis

P. Esponda1 and R. Carballada

Centro de Investigaciones Biológicas. CSIC. Madrid. Spain.

1 To whom correspondence should be addressed, Dr. Pedro Esponda. Centro de Investigaciones Biológicas (C.S.I.C.), Ramiro de Maeztu 9. 28040 Madrid. Spain. Telf: 3491 8373112, Fax: 3491 5360432, E. Mail: esponda{at}cib.csic.es

Mouse cauda epididymis were in-vivo transfected using the lipid FuGENE 6 as gene vector. Two gene constructions were employed: the p-GeneGRIP which codifies for the GFP (Green Fluorescent Protein) and the pSEAP-control that expresses an Alkaline Phosphatase as a secretion. Transfection was detected by fluorescence and appeared in the nucleus and cytoplasm of epithelial cells. Transfection was observed in 39.70% of cells after 2 days and in 31.77 % after 7 days, and then diminished progressively. Moreover, the presence of the transgene in the DNA isolated from treated epididymides was observed by PCR. GFP gene expression appeared in large areas of the cauda epididymis and it was observed exclusively in the cytoplasm of epithelial cells. GFP gene expression occurred during two weeks after gene injection and occupied 32.24%, 29.98% and 22.37% of the area of the tubules when analyzed 2, 7 and 15 days after gene injection. The cauda was also analyzed in toto and showed similar results. The use of the pSEAP-control gene showed that cauda epididymis secretions can also be modified by the transfection procedure. A significant increase of alkaline phosphatase activity appeared in the epididymal fluids seven days after gene injection. These results indicate that transfection procedures could be an important tool in the future to study epididymal physiology or to change the fertilizing ability of spermatozoa.

Key Words: Epididymis/Gene transfer/Lipid Vectors/Transfection

Submitted on July 18, 2009; resubmitted on March 25, 2009; accepted on March 27, 2009.


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