Sequestration of PDC-109 protein improves freezability of crossbred bull spermatozoa

Abstract

A study was carried out to assess the effect of sequestration of PDC-109 protein, a majority constituent of heparin binding proteins (HBP) of seminal plasma, on freezability and in vitro fertilizing ability of crossbred bull spermatozoa after cryopreservation. The study consisted of isolation and characterization of PDC-109 protein to raise anti-sera against it in rabbits. Following which, raised antibodies against PDC-109 protein was quantitated and coated in tubes used for collection of ejaculates. Semen ejaculates thus collected were cryopreserved using EYTG extender. Physico-morphological characteristics, viz. motility, viability, acrosomal integrity and HOS response as an indicator of freezability of cryopreserved spermatozoa were determined at pre freeze as well as post thaw stage. At pre freeze stage, a significant (p<0.05) improvement in viability (83.83±2.18 vs 75.17±2.42) and acrosome integrity (81.33±2.38 vs 72.83±2.39) in antibodies treated group than control was observed. Similarly, increase in HOS responsive spermatozoa was highly significant (p<0.01) than control (78.83±1.69 vs 67.5±1.75). At post thaw stage, significant (p<0.05) improvement in viability (69.50±2.16 vs 60.33±2.19) and HOS responsive spermatozoa (68.67±1.62 vs 58.50±1.32) and highly significant (p<0.01) increase in individual motility (56.17±1.83 vs 47.00±1.86) and acrosome integrity (75.17±2.38 vs 61.83±2.1) was observed in antibodies treated group when compared to control was observed. The results from the study revealed that sequestration of PDC-109 protein from semen samples leads to significant improvement in pre-freeze and post-thaw values of above parameters in cryopreserved spermatozoa. It is thus concluded that sequestration of PDC-109 protein from ejaculates improves freezability of crossbred bull spermatozoa.