title: Effect of cryopreservation on phosphorylation state of proteins involved in sperm motility initiation in sea bream.
abstract: We have previously demonstrated that in sea bream Sparus aurata motility initiation determined changes in the phosphorylation state of some proteins. This paper describes an investigation of the effect of a freezing-thawing procedure on the protein phosphorylation/dephosphorylation pattern. Proteins extracted from fresh and cryopreserved spermatozoa (before and after motility activation) were separated on SDS-PAGE, blotted on nitrocellulose membrane and treated with anti-phosphotyrosine, anti-phosphothreonine, or anti-phosphoserine antibodies. The results obtained demonstrate that the cryopreservation protocol has a strong effect on the phosphorylation state of proteins. In general, compared to fresh sperm, phosphorylated proteins are most numerous in both activated and non-activated cryopreserved sperm, and in particular we observed a dramatic increase in threonine phosphorylation. However, frozen-thawed sperm showed a minor number of proteins that changed their phosphorylation state after motility activation. Among these, we identified the acetyl-coenzyme A synthetase that plays a role in sperm motility initiation in both fresh and cryopreserved sperm.
authors.affiliation: Department of Biological and Environmental Sciences and Technologies, Laboratory of Comparative Physiology, University of Salento, Via Provinciale Lecce-Monteroni, 73100 Lecce, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Zilli
authors.forename: R
authors.initials: R
authors.lastname: Schiavone
authors.forename: C
authors.initials: C
authors.lastname: Storelli
authors.forename: S
authors.initials: S
authors.lastname: Vilella
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D021541:Sea Bream / Q000378:metabolism*; D012662:Semen Preservation; D013081:Sperm Motility / Q000502:physiology*; D013094:Spermatozoa / Q000378:metabolism
chemical_list: D011506:Proteins