Expression of HSP90 Gene in the Cryopreserved Bovine Spermatozoa
Keywords:
cryopreservation, HSP90, heat shock, spermatozoa, bullsAbstract
Cryopreservation is a technique, commonly used for a long-term storage of male gametes as an effective way to
preserve their longevity and vitality. During cryopreservation and following the thawing process, sperm cells are
exposed to thermal stress, which may lead into disruptions of their morphological structures and a subsequent
functional dysfunction. The objective of our study was to monitor HSP90 (heat shock protein 90) gene expression in
cryopreserved bovine spermatozoa. HSP90 is one of the main heat-related cellular proteins, which belongs into
chaperone protein family. It is involved in the formation of protein complexes, which stabilize proteins against heat
stress and aid in the protein degradation. For the experimental purposes were used semen samples from 12 sexually
mature Holstein-Friesian bulls. The samples were divided into two identical fractions, one fraction was
cryopreserved and kept in liquid nitrogen at -196°C, while the other fraction was processed as non-cryopreserved
fresh semen. The sperm motility rate was evaluated using CASA (Computer Assisted Semen Analysis).
Quantification of isolated RNA was performed by real-time PCR and the expression of the HSP90 gene was assessed
with the Western blot technique. For a proper analysis, we compared cryopreserved sperm samples with fresh ones.
The data from our study indicate a reduction in the sperm motility (P<0.01) and HSP90 gene expression by
approximetly 50% in the cryopreserved samples (P<0.01 for real-time PCR; P<0.05 for Western blot). Based on the
results, we may confirm that the gene expression is reduced in the cryopreserved samples, which leads into a
decrease of the protective properties of HSP90 due to the impact of low temperatures.
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