CORRELATION BETWEEN SPERM MEMBRANE DESTABILIZATION BY HEPARIN AND ANILINE BLUE STAINING AS MEMBRANE INTEGRITY INDEX

Acidic aniline blue stain (AAB) was studied in relation to sperm membr ane destabilization and nuclei decondensation by heparin. Untreated sp ermatozoa smears stained with AAB or vital stain shows 28.4% of staine d and 71.6% of unstained nuclei. This behavior was also observed when incubation was done in the presence of 5 mM glutathione (GSH) used alo ne. In the presence of 21.6 mu M heparin, staining of sperm cells comm enced 10 min after heparin addition and was dependent on the incubatio n time. During the experiment 12.3% of the total cholesterol content a nd 20 mu g protein/10(8) sperm cells were released. In the presence of 21.6 mu M heparin-5 mM/GSH, swelling of sperm nuclei reach 95% after 150 min incubation. When this experiment was Nn along with AAB, the sa me average (45%) was seen in the first 30 min, which gives plenty of t ime to trigger the nuclei's decondensation mechanism. The percentage o f stained cells was of 71%, indicating that the histone is not complet ely replaced, and insuring a positive reaction with AAB stain. It woul d appear that AAB stain can be used as a membrane integrity index to c onfirm the destabilization effect of heparin on the sperm membrane.