Abstract
Heat Shock Protein 70 (HSP70) induction helps alleviate the adverse effects of several types of stress
in the cells. It can also play a significant role in sperm protection and survival during the extreme
conditions of the semen cryopreservation process. The present study was conducted to investigate the
role of this protein in spermatozoa of water buffalo by analyzing the changes in its gene expression,
protein induction, and distribution after cryopreservation. Differences in HSP70 mRNA and protein
levels between fresh and frozen semen samples were determined through real-time PCR and western
blot analysis, respectively. Cellular localization patterns of HSP70 in the different sample groups were
then detected in an indirect immunofluorescence assay. Lastly, the average sperm motilities of the
semen sample groups were evaluated and compared. Results showed that the HSP70 mRNA expression
decreased by fifty percent (P<0.05) while the protein level significantly rose by sixty percent (P<0.05)
in the frozen sample group. The localization of HSP70 protein in the sperm head also shifted from the
equatorial segment in the fresh sample group to the post-acrosomal region in frozen semen samples.
Moreover, the average sperm motility appeared to fluctuate with the mRNA level and comparably
decreased by fifty percent (P<0.05) in the frozen sample group. These changes on the HSP70 in
spermatozoa exhibited the adverse effects of the underlying capacitation-like event in the
cryopreservation process. Overall, these findings presented a valuable insight into the current buffalo
semen cryopreservation technique and a potential approach to its improvement.
Maylem, E.R.S, Rivera, S.M, Ramos, G.E, Atabay, E.C, Venturina, E. V, Atabay, E.P, Shanemae M. Rivera. (2021) Changes on the Heat Shock Protein 70 (HSP70) in Water Buffalo Spermatozoa revealed the Capacitation-like event in Cryopreservation, Asian Journal of Agriculture and Biology, Volume-09, Issue-2.
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