In vivo electroporation of the testis versus transgenic mice model in functional studies of spermatocyte‐specific hst70 gene promoter: A comparative study

Abstract

To determine whether DNA transfer to mouse testes by in vivo electroporation could be useful method for studying regulatory elements of genes specifically active in spermatocytes first we compared the expression pattern of a construct containing the EGFP reporter gene ligated to a fragment of the heat shock testis-specific hst70 gene promoter, both in testis of transgenic mice and in testis electroporated in vivo. While in transgenic mice the EGFP was expressed in all seminiferous tubules in a cell- and stage-specific manner, in the testes electroporated in vivo only small fraction of cells expressed this marker protein. In order to make a quantitative comparison between the specificity of these two experimental systems we used several vectors containing the CAT gene ligated to fragments of the hst70 gene 5' upstream of DNA sequences which either promoted or did not activate expression of the reporter gene in the testes of transgenic mice. Also, as a reference opposite to spermatogenic cells we examined the expression pattern of the same set of vectors in the rat hepatoma FTO 2B cells. Although electroporated testes retain some spermatocyte-specific features such as the ability to repress promoters which do not contain regulatory elements responsible for testis-specific transcription, several important drawbacks of the method are evident. They include basal activity of constructs which are not transcribed in testes of transgenic mice and low overall transfection efficiency. This may hamper studies in which subtle changes in the expression pattern are under investigation. However, the in vivo electroporation of the testis can be useful for preliminary screening of constructs aimed to study in transgenic mice.

Yin Yang-1 Regulates the Characterized Murine Focal Adhesion-Associated Protein Promoter

“…In vivo electroporation is an effective tool for promoter analysis that have been accomplished in muscle (Bertrand et al, 2003), testis (Widlak et al, 2003;Somboonthum, 2005), and epididymis (Kirby et al, 2004). Differences in our in vivo results from testis electroporation and in vitro results from cell transfection indicated that activities of the FAAP promoter constructs were substantially variable in different conditions.…”

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

“…In vivo gene transfer into the testis by electroporation offers a potential alternative to standard transgenic approaches for studying testis and sperm function and as a novel way of generating transgenic animals (12)(13)(14)(15)(16)(17)(18)(19)25). However, previous studies using this approach have failed to show whether it is possible to use this technique to study the pattern of localization of a sperm protein of physiological interest, as opposed to simply a reporter protein.…”

Duration and level of transgene expression after gene electrotransfer to skin in mice

“…[1][2][3] A vast number of studies have encompassed gene electrotransfer to muscles, [4][5][6][7][8] but the technique is also efficient in, for example, cornea, 9 lungs, 10 liver 11,12 kidney, 13 bladder, 14 testis, 15 skin 16,17 and tumor. [18][19][20] One very interesting perspective with gene electrotransfer is that the choice of tissue can determine the duration of expression of the transgene.…”