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Sequestration of PDC ‐109 Protein by Specific Antibodies and Egg Yolk Cryoprotects Bull Spermatozoa
Author(s) -
Srivastava N,
Srivastava SK,
Ghosh SK,
Jerome A,
Das GK,
Mehrotra S
Publication year - 2013
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.12151
Subject(s) - extender , cryopreservation , andrology , yolk , sperm , chemistry , semen , antibody , acrosome , biology , immunology , embryo , food science , microbiology and biotechnology , medicine , organic chemistry , polyurethane
Contents PDC ‐109, one of the most abundant proteins in bovine seminal plasma, has detrimental effect on spermatozoa in a time‐ and concentration‐dependent manner. Therefore, we hypothesized that sequestration of detrimental protein from ejaculates would be beneficial following cryopreservation of sperm cells. To this aim, we evaluated the effect of sequestration of PDC ‐109 either by anti‐ PDC ‐109 antibodies ( A b) or egg yolk ( EY ) alone or by the synergistic action of EY + A b in minimizing cryoinjury to bull spermatozoa. PDC ‐109 protein was purified by applying two‐step chromatography procedures. The purified protein was injected in rabbits to raise antibodies which were isolated using ion‐exchange chromatography. After checking the A b cross‐reactivity, they were quantitated and added to ejaculates, either alone or in addition to EY in Tris‐glycerol ( TG ) extender. Thus, ejaculates were processed in extender containing EY + TG (group I), A b + TG (group II) or EY + A b + TG (group III). Semen quality parameters ( SQP s) viz. viability and acrosome integrity ( FITC ‐ PSA ), cryoinjury to spermatozoa (chlortetracycline, CTC assay) and in vitro fertility of protein‐sequestered‐semen (zona‐penetration assay) were evaluated. A significant (p < 0.05) improvement in post‐thaw SQP s as well as in non‐capacitated spermatozoa observed at pre‐freeze and post‐thaw stages of cryopreservation in group III compared with other groups indicated reduction in protein‐mediated cryoinjury. From this study, it can be concluded that sequestration of PDC ‐109 by synergistic action of EY + A b as compared to either of them alone significantly improve sperm quality and minimize cryoinjury to bull spermatozoa upon storage at ultra‐low temperatures.