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Abstract
Several reliable methods to produce transgenic animals use the male genome. After penetration into oocytes, sperm DNA undergoes dramatic conformational changes that might represent an opportunity for exogenous DNA to integrate into the zygote genome. A nuclease, DNase I, with Ca 2+ /Mg 2+ dependent activity and Zn 2+ inhibition, is one of the enzymes responsible for sperm DNA remodeling. To date, the effect of different calcium concentrations in manipulation media on porcine intracy- toplasmic sperm injection has not been fully investigated. The present study was conducted to examine the effect of calcium in the surrounding media, and we found that the number of embryos expressing green fluorescent protein (EGFP) was increased in media containing Ca 2+ . How- ever, the number did not change over Ca 2+ concentrations from 2 to 8 mmol$L -1 (P>0.05). Moreover, free Ca 2+ concentrations in the media were found to affect the efficiency which is porcine intracytoplasmic sperm injec- tion (ICSI) embryos expressing EGFP protein, which was related to the activation of ooplasmic DNase I. These findings reveal a mechanism and pathway involving EGFP expression in ICSI embryos.
Highlights
Transgenesis represents an important tool in basic research as well as in livestock production
The results (Table 1) for the group without electrical activation demonstrated that the rates of cleavage varied with concentrations of calcium added to the media
The number of embryos expressing expressing green fluorescent protein (EGFP) was significantly increased in the media containing Ca2+, whereas the number did not change with Ca2+ concentrations from 2 to 8 mmol$L–1 (P > 0.05)
Summary
Transgenesis represents an important tool in basic research as well as in livestock production. Several reliable methods to produce transgenic animals use the male genome for exogenous DNA integration. Male pronuclear injection is currently the most frequently used method to generate transgenic animals.
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