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

Characterization of two heterozygous mutations of the oocyte activation factor phospholipase C zeta (PLCζ) from an infertile man by use of minisequencing of individual sperm and expression in somatic cells

“…It is also worth noting that Ito et al (26) demonstrated nuclear localization of human and mouse PLCz in COS-7 cells. However, Coward et al (27) found that, in COS cells, mouse PLCz was localized to either the nucleus or cytoplasm. Furthermore, Kashir et al (20) indicated a reticular cytoplasmic pattern of localization for human PLCz within HEK293T cells, possibly indicating cytoplasmic organelle(s).…”

Phospholipase C zeta undergoes dynamic changes in its pattern of localization in sperm during capacitation and the acrosome reaction

“…Full-length mouse PLCz cDNA was isolated by PCR from a construct in which the full-length mouse PLCz had been linked to EYFP (24) and ligated into the pGEX2TKP GST fusion protein expression vector (GE Healthcare Life Sciences, Amersham, UK). The construct was verified by DNA sequencing (MWG Biotech).…”

Future Perspectives in the Diagnosis and Management of Unexplained Male Infertility

“…In vivo electroporation is another technique that involves injecting a DNA construct into the testes with application of electric pulses that permeates cell membranes and allow DNA to enter the cells. Using in vivo electroporation, groups have demonstrated the ability to express transgenes in mature epididymal sperm in mice [49] and hamsters [50]. They also demonstrated that the use of electroporation in vivo does not lead to adverse effects on testicular integrity and sperm quality [49].…”

“…Using in vivo electroporation, groups have demonstrated the ability to express transgenes in mature epididymal sperm in mice [49] and hamsters [50]. They also demonstrated that the use of electroporation in vivo does not lead to adverse effects on testicular integrity and sperm quality [49]. However, the expression of these transgenes is typically transient and may last as long as 60 days.…”