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Cryopreservation of common carp (Cyprinus carpio L.) sperm induces protein phosphorylation in tyrosine and threonine residues
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SYSNO ASEP 0425785 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Cryopreservation of common carp (Cyprinus carpio L.) sperm induces protein phosphorylation in tyrosine and threonine residues Author(s) Li, P. (CZ)
Hulák, M. (CZ)
Li, Z. - H. (CZ)
Šulc, Miroslav (MBU-M) RID, ORCID
Pšenička, M. (CZ)
Rodina, M. (CZ)
Gela, D. (CZ)
Linhart, O. (CZ)Source Title Theriogenelogy. - : Elsevier - ISSN 0093-691X
Roč. 80, č. 2 (2013), s. 84-89Number of pages 6 s. Language eng - English Country US - United States Keywords Cryopreservation ; Sperm ; Phosphorylation Subject RIV CE - Biochemistry Institutional support MBU-M - RVO:61388971 UT WOS 000320742600003 DOI 10.1016/j.theriogenology.2013.03.021 Annotation The effect of cryopreservation on the protein phosphorylation/dephosphorylation pattern of common carp (Cyprinus carpio) sperm is described. Sperm was diluted in dimethyl sulfoxide (DMSO) and ethylene glycol (EG)-based extenders, followed by equilibration, freezing, and thawing. Proteins extracted from fresh and cryopreserved spermatozoa were separated on SDS-PAGE and two-dimensional gel electrophoresis, blotted on polyvinylidene difluoride membrane, and treated with anti-phosphotyrosine, anti-phosphothreonine, or anti-phosphoserine antibodies. For the subsequent protein identification we used matrix-associated laser desorption/ionization time-of-flight mass spectrometry. The results demonstrated that cryopreservation with either DMSO or EG extender significantly altered the phosphorylation state of sperm proteins on tyrosine or threonine residues. A dramatic decrease in tyrosine phosphorylation was detected in the cryopreservation procedures with DMSO extender. Endoplasmin, transketolase, and S-adenosylhomocysteine hydrolase were identified as proteins that play a key role in cellular stress responses and oxidation and/or reduction reactions. Results indicate that the phosphorylation and/or dephosphorylation modifications of sperm proteins that occur during cryopreservation could stimulate a series of biochemical effects interfering with spermatozoa function and leading to a loss of motility and fertilization ability. Our findings indicated that use of EG extender provided superior protein preservation during sperm storage Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2014
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