CHARACTERIZATION OF MALABARI BUCK SEMINAL PLASMA PROTEINS IN RELATION TO SEMEN FREEZABILITY AND FERTILITY
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Date
2016-12-30
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COLLEGE OF VETERINARY AND ANIMAL SCIENCESMANNUTHY, THRISSUR
Abstract
The study was undertaken to characterize the protein profile of Malabari buck seminal
plasma for identifying protein markers of freezability and fertility and to compare the in vivo
fertility of Malabari buck semen with low and high semen freezability. Adult Malabari bucks
with post thaw sperm progressive motility of more than 35 per cent were classified as having
high semen freezability while those with less than 30 per cent were classified as having low
semen freezability (four each) were utilized for the study.
Tris - egg yolk based extender was used for cryopreservation of the ejaculates (n = 48; 24
each from bucks having high and low semen freezability). Significantly higher values were
noted in the pre-freeze acrosome integrity and membrane cholesterol levels and post-thaw
motility, viability, acrosome integrity, hypo osmotic sperm swelling response (HOS) and
membrane cholesterol levels of high semen freezability bucks when compared with low semen
freezability bucks. Whereas the seminal plasma ALP and LDH were found to be significantly
lower in high semen freezability bucks compared with low semen freezability bucks at post
thaw stage.
The total protein content and average number of protein bands in seminal plasma of high
and low semen freezability bucks did not differ significantly (75.82 ± 4.62 mg/ml vs 67.07 ±
6.07 mg/ml; 20.83 ± 0.53 vs 22.09 ± 0.49). The 24 kDa, 79 kDa, 84 kDa and 134 kDa proteins
were found to have significantly higher occurence in seminal plasma of bucks with high semen
freezability whereas the occurence of 15 kDa, 19.9 kDa, 21.5 kDa, 70kDa and 89 kDa proteins
were found significantly higher in seminal plasma of bucks with low semen freezability. On
analysis of 2D gels, an average of 67.67 ± 4.06 and 75.33 ± 4.28 spots (pI 3-10) were detected
in semen of high and low semen freezability bucks, respectively and the difference was not
significant. On western blot analysis, osteopontin, a protein fertility marker, was found to be
present in seminal plasma of all the bucks studied.
For fertility studies 134 does were inseminated with semen preserved by chilling and 91
with cryopreserved semen. The conception rate (CR) with cryopreserved semen was
significantly higher in bucks with high semen freezability than with low semen freezability. In
the semen preserved by chilling, no significant difference was noticed between low and high
semen freezability animals in the spermatozoa progressive motility, viability, acrosome
integrity, HOS response or seminal plasma ALP and LDH levels after dilution or after 24 h of
storage at refrigeration. But when chilled semen was used, the CR was significantly higher for
bucks with low semen freezability than for bucks with high semen freezability.
Description
Thesis Submitted in partial fulfilment of the requirement for the degree of Doctor of Philosophy in Animal Reproduction, Gynaecology and Obstetrics