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Statements

Subject Item
n2:RIV%2F61388971%3A_____%2F13%3A00425785%21RIV14-AV0-61388971
rdf:type
n12:Vysledek skos:Concept
dcterms:description
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 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
dcterms:title
Cryopreservation of common carp (Cyprinus carpio L.) sperm induces protein phosphorylation in tyrosine and threonine residues Cryopreservation of common carp (Cyprinus carpio L.) sperm induces protein phosphorylation in tyrosine and threonine residues
skos:prefLabel
Cryopreservation of common carp (Cyprinus carpio L.) sperm induces protein phosphorylation in tyrosine and threonine residues Cryopreservation of common carp (Cyprinus carpio L.) sperm induces protein phosphorylation in tyrosine and threonine residues
skos:notation
RIV/61388971:_____/13:00425785!RIV14-AV0-61388971
n12:predkladatel
n15:ico%3A61388971
n4:aktivita
n16:S n16:P n16:I
n4:aktivity
I, P(ED2.1.00/01.0024), S
n4:cisloPeriodika
2
n4:dodaniDat
n9:2014
n4:domaciTvurceVysledku
n18:9501894
n4:druhVysledku
n13:J
n4:duvernostUdaju
n19:S
n4:entitaPredkladatele
n10:predkladatel
n4:idSjednocenehoVysledku
67369
n4:idVysledku
RIV/61388971:_____/13:00425785
n4:jazykVysledku
n14:eng
n4:klicovaSlova
Cryopreservation; Sperm; Phosphorylation
n4:klicoveSlovo
n6:Sperm n6:Phosphorylation n6:Cryopreservation
n4:kodStatuVydavatele
US - Spojené státy americké
n4:kontrolniKodProRIV
[043301E5B434]
n4:nazevZdroje
Theriogenelogy
n4:obor
n7:CE
n4:pocetDomacichTvurcuVysledku
1
n4:pocetTvurcuVysledku
8
n4:projekt
n17:ED2.1.00%2F01.0024
n4:rokUplatneniVysledku
n9:2013
n4:svazekPeriodika
80
n4:tvurceVysledku
Linhart, O. Rodina, M. Pšenička, M. Li, P. Gela, D. Hulák, M. Li, Z. - H. Šulc, Miroslav
n4:wos
000320742600003
s:issn
0093-691X
s:numberOfPages
6
n5:doi
10.1016/j.theriogenology.2013.03.021