DETECTION OF THE Y-CHROMOSOME IN BOVINE SPERMATOZOA BY RADIOACTIVE INSITU HYBRIDIZATION

The in situ hybridization with radioactive labelled Y-specific DNA probes was used for the quantification of X- and Y-chromosome bearing sperm. The hybridization of female and male bovine lymphocyte's standard mixture indicates, that the proportion of both cell types can be detected on the basis of hybridization signals. The efficient decondensation with trypsin and DTT revealed appropriately 49.3% labelled spermatozoa among 3,000 scored, which is not different from the theoretical. The applicability of the in situ hybridization to monitor the success of sperm separation methods was demonstrated at the example of BSA gradient and Percoll centrifugation.