Production Of Transgenic Rabbit Embryo By Intracytoplasmic Sperm Injection

Studies were carried out to find out if rabbit spermatozoa can act as an effective vector for carrying exogenous DNA during the production of transgenic rabbit embryo by intracytoplasmic sperm injection. The capacity of sperm to bind exogenous DNA and factors affecting this process were analyzed. In this study green fluorescent protein gene was selected as a vital report during production of transgenic rabbit embryo by intracytoplasmic sperm injection.Fully capacitated rabbit sperm could bind exogenous tetramethylrhodamine-6-dUTP labeled DNA. DNA absorbtion by sperm cells increased following the times of washing but the vitality of the sperm was coming down after more times of washing.The rabbit sperm frozen in liquid nitrogen absorbed more exogenous DNA and resuilts were more reproducable than washed sperm. It was found that some DNA was absorbed in the surface of spermatozoa but in some case DNA was incorporated into the sperm.Sperm of Epididymides and ejaculation could bind tetramethylrhodamine-6-dUTP labeled exogeous DNA. The ability of binding exogenous DNA by sperm of epididymis and ejaculation were no difference statistically. Using Sperm of Epididymis and ejaculation in the production of embryos by intracytoplasmic sperm injection it was found that the fertilization rate (21.6%and 23%)and positive rate (15.5% and 16.5%) were very close.In the experiments of production of transgenic rabbit embryos by intracytoplasmicing sperm injection, the cleavage rate was low ( 15.5%) but positive rate was high (28.6% - 44%) among the 2-16 cell embryos.