Evaluation of fertilizing capacity of rabbit sperm on the basis of annexin v-labelled membrane changes

Abstract

The aim of this study was (1) to analyze fresh intact or differently influenced rabbit sperm samples, using Annexin V–binding method for the detection of membrane phosphatidylserine translocation, and (2) to estimate obtained results in relation to the conceptional rate of rabbit females fertilized with semen from different groups. Following sample groups were designed: 1. freshly collected semen; 2. semen after 90 min incubation in sperm diluent; 3. semen incubated in cryoprotective medium, without freezing; 4. semen frozen in floating chamber with liquid nitrogen; 5. Semen frozen using classical procedure in manual regime. Higher rate of annexin V-positive (AnV+) cells was observed in both cryopreserved groups (4, 5) when compared with groups 1, 2 and 3 (p< 0.05). The highest conceptional rate of rabbit females was obtained, when semen doses from groups 3, 2 or 1 were used for the insemination. Oppositely, conceptional rate was minimal when sperm was frozen in floating chamber (group 4; 16.67%), whilst using the sperm frozen in manual regime (group 5) no one of 13 inseminated females was parturited. A negative linear dependence between the rate of An-V+ cells and female conceptional rate was found. Therefore, Annexin V-labelling would be considered as a technique of choice to predict fertilizing capacity of rabbit sperm.