Abstract:

Objective

To compare the influence of three different puncture sites of in utero electroporation on the death rate of mice embryos,the thickness and the area of cortex,cell differentiation,cell proliferation,cell migration and cell apoptosis.

Methods

It groups on the bases of common puncture sites as follows:1.electric shock only(control);2.injection near the posterior fontanel,at about one millimeter(here,group A);3.injection along the antero-posterior axis into the cephalic ventricle(group B);and 4.injection near the midpoint between the anterior fontanel and posterior fontanel,at about three millimeters(group C).We compared the four groups using littermate mice,and repeated the experiment with 5 pregnant mice.To detect whether different injection positions affect cell differentiation,cell proliferation,cell migration and cell apoptosis,we stained cells with anti-Tbr1,anti-Tbr2,anti-Pax6 anti-Ctip2,anti-Caspase-3 and anti-Ki67.In differentiation,proliferation,and apoptosis assays,the cell number was analyzed with SPSS 13.0 software,using one way ANOVA.

Results

We found no statistical significant differences between the three puncture methods in the death rate of embryos,the thickness and the area of cortex,cell differentiation,cell proliferation,cell migration and cell apoptosis(P>0.05).

Conclusion

Three puncture sites used for in utero electroporation show no significantly different negative impacts during gene transfer into the embryonic mousebrain.

Key words: Genetransfer techniques, Puncture, Cell differentiation, Cell proliferation