Journal
CELL AND TISSUE RESEARCH
Volume 371, Issue 2, Pages 365-373Publisher
SPRINGER
DOI: 10.1007/s00441-017-2705-x
Keywords
Sperm maturation; Protein phosphorylation; Epididymis; Bull sperm; Anti-phosphotyrosine antibodies
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Funding
- Scientific Grant Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic
- Slovak Academy of Sciences [VEGA-2/0037/16]
- Slovak Research and Development Agency [APVV-15-0196]
- bilateral project SAS-CAS [15-05]
- CIGA [20162027]
- project BIOCEV - Biotechnology and Biomedicine Centre of the Academy of Sciences
- Charles University [CZ.1.05/1.1.00/02.0109]
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Phosphorylation, or dephosphorylation, is one of the most frequent post-translational modifications regulating protein-protein activity in eukaryotic cells. Whereas mature spermatozoa (as specialized cells) are transcriptionally inactive and do not synthesize new proteins, phosphorylation of sperm proteins is very important for the regulation of the sperm function. Although the post-testicular maturation of spermatozoa is a process common to all mammals, comparative studies showed significant differences in sperm surface proteins and the mechanisms of protein modification during the epididymal maturation. In our study, the evaluation of tyrosine phosphorylation, represented by the fluorescent patterns of used anti-phosphotyrosine antibodies (P-Tyr-01 and 4G10), in spermatozoa isolated from different regions of the epididymis - caput, corpus and cauda - was performed. Although in general both antibodies detected almost the same reaction patterns, we observed some dissimilarity associated with the binding specificity of the antibodies and also the segment-dependent manner of phosphorylated protein localization. These data were filled up by immunohistochemical analysis of testes and epididymides cryosections. Additionally, our phosphoproteomic study focused on evaluation of the changes in the pattern of tyrosine-phosphorylated proteins during the post-testicular maturation of bull spermatozoa (PY20 antibody). To summarize the results, an increasing trend of tyrosine phosphorylation of proteins during the maturation of bull sperm in the epididymis was consistently observed in all the methods/experiments.
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