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Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

100 PROTEOME OF BULL SPERMATOZOA

J. M. Feugang, C. Rozanas, A. Kaya, E. Topper and E. Memili

Reproduction, Fertility and Development 20(1) 130 - 130
Published: 12 December 2007

Abstract

The reproductive performance of a herd is the biggest factor affecting production and product quality of livestock. Thus, a decline of male fertility represents a dramatic economic loss in beef and dairy industries. Caused by molecular defects in the spermatozoa, uncompensatory infertility is a current challenge for the cattle industry, because even with normal sperm morphology, motility, and number, fertility of bulls is still sub-par. Thus, the objective of this study was to determine the global proteome of spermatozoa collected from bulls with different fertility and study the proteins playing a role in uncompensatory infertility. We performed difference gel electrophoresis (2D-DIGE) using cryopreserved sperm from a total of 6 bulls. Spermatozoa were thawed, purified by percoll gradient, and washed in PBS solution. For each bull, a pellet of 100 million sperm cells was resuspended in the 2D-DIGE labeling buffer, and the total protein was quantified. Cell lysates were separately labeled with CyDye DIGE Fluor dyes (and reciprocally with different dyes) and multiplexed in pairs on 3 gels. The samples were focused according to their isoelectric point through an Immobiline DryStrip, pH4-7, using a IPGphor 2, followed by separation on 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels (21.5–200 kDa) using a DALTsix. The resulting gels were scanned, and their topographic digital maps were used for algorithmic spot matching, background normalization, spot differences quantification, and elimination of artifacts (DeCyder 2-D Differential Analysis Software, GE Healthcare Life Sciences, Piscataway, NJ, USA). The results showed between 2600 and 2800 proteins with high confidence. Compared to the high-fertile bulls, 30 proteins were increased, and 27 were decreased in the low-fertility bulls within a 2-fold range. The largest significant increase and decrease were 3.97- and 2.4-fold, respectively. The identification of these differentially represented proteins is in progress. However, our results provided a panoramic view of sperm proteome from bulls of different fertility and thus paved the way for research on mechanisms of uncompensatory bull infertility.

https://doi.org/10.1071/RDv20n1Ab100

© CSIRO 2007

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