title: Identification of differentially expressed proteins in fresh and frozen-thawed boar spermatozoa by iTRAQ-coupled 2D LC-MS/MS.
abstract: Cryodamage is a major problem in semen cryopreservation, causing changes in the levels of proteins that influence the function and motility of spermatozoa. In this study, protein samples prepared from fresh and frozen-thawed boar spermatozoa were compared using the isobaric tags for relative and absolute quantification (iTRAQ) labeling technique coupled to 2D LC-MS/MS analysis. A total of 41 differentially expressed proteins were identified and quantified, including 35 proteins that were present at higher levels and six proteins that were present at lower levels in frozen-thawed spermatozoa by at least a mean of 1.79-fold (P<0.05). On classifying into ten distinct categories using bioinformatic analysis, most of the 41 differentially expressed proteins were found to be closely relevant to sperm premature capacitation, adhesions, energy supply, and sperm-oocyte binding and fusion. The expression of four of these proteins, SOD1, TPI1, ODF2, and AKAP3, was verified by western blot analysis. We propose that alterations in these identified proteins affect the quality of cryopreserved semen and ultimately lower its fertilizing capacity. This is the first study to compare protein levels in fresh and frozen-thawed spermatozoa using the iTRAQ technology. Our preliminary results provide an overview of the molecular mechanisms of cryodamage in frozen-thawed spermatozoa and theoretical guidance to improve the cryopreservation of boar semen.
authors.affiliation: The Key Laboratory for Farm Animal Genetic Resources and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Chen
authors.forename: Huabin
authors.initials: H
authors.lastname: Zhu
authors.forename: Chuanhuo
authors.initials: C
authors.lastname: Hu
authors.forename: Haisheng
authors.initials: H
authors.lastname: Hao
authors.forename: Junfang
authors.initials: J
authors.lastname: Zhang
authors.forename: Kunpeng
authors.initials: K
authors.lastname: Li
authors.forename: Xueming
authors.initials: X
authors.lastname: Zhao
authors.forename: Tong
authors.initials: T
authors.lastname: Qin
authors.forename: Kan
authors.initials: K
authors.lastname: Zhao
authors.forename: Huishan
authors.initials: H
authors.lastname: Zhu
authors.forename: Dong
authors.initials: D
authors.lastname: Wang
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000032:analysis / Q000378:metabolism; D002853:Chromatography, Liquid; D015925:Cryopreservation / Q000662:veterinary; D005615:Freezing / Q000009:adverse effects; D008297:Male; D018384:Oxidative Stress; D055101:Semen Analysis / Q000379:methods; D012662:Semen Preservation / Q000009:adverse effects / Q000662:veterinary; D029589:Seminal Plasma Proteins / Q000032:analysis* / Q000378:metabolism; D013094:Spermatozoa / Q000737:chemistry* / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*; D013552:Swine / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000975:Antioxidants; D029589:Seminal Plasma Proteins; C425042:spermadhesin