The Expression And Application Of ZFY And TSPY In Holstein Cattle Sperm

The ability to pre-select offspring sex via separation of X and Ybearing sperm would have profound ramifications for the animal husbandry industry.Commercially available flow sorting is another way for sex sorting.However,its wide used is restricted because of its high price and effects on DNA damage.In addition that this will cause DNA damage in sperm.Identification of a specific cell surface marker for immune selection will bring a major advance in these fields by a)allowing for batch selection and b)reducing the safety issues associated with DNA damage detection methods currently used.Therefore,this study is aimed to find differential proteins between X and Y sperms and isolate them by researching TSPY and ZFY.We apply following procedure:(1)Localization and expression of these genes were performed in Holstein sperm.(2)Protein expression characteristic in Holstein sperm and sex controlled sperm was analyzed.(3)Finally we sorted Holstein sex controlled sperm by using antibodies of these two genes which enabled us to confirm whether or not these two genes are differential expression in X/Y sperm and whether they could be used.The main results of this study are as follows:1.Our result revealed that the ZFY protein was mainly located at tail of the sperm with weak signal but TSPY protein located at acrosome and nucleus with positive signals in sperm.2.Expression of ZFY and TSPY in sorted Y sperm were both significantly higher than sorted X sperm(P<0.01).The positive rates of antigen-ZFY antibodies immune response in sorted Y sperm(35.51±0.79%)was significantly higher(P<0.01)than sorted X sperm(13.47±0.18%).The positive rates of antigen-TSPY antibodies immune response of sorted Ysperm(41.75±4.05%)was remarkable higher(P<0.01)than sorted X sperm(19.30±1.32%).3.Sorting sperm by using anti-ZFY and anti-TSPY antibodies with flow cytometry separation technology:Our result show that sorted positive antigen-ZFY antibodies sperm up to 77.7%,and sorted positive antigen-ZFY antibodies sperm up to 82.2%.In summary,TSPY protein located at the acrosome and nucleus of Y sperm but ZFY protein was located at tail of Y sperm.Also we concluded that these genes are similar and have potential application value in sorting X/Y sperm.