Profiling of differentially expressed proteins between fresh and frozen-thawed Duroc boar semen using ProteinChip CM10

Yong-Min Kim; Sung-Woo Park; Mi-Jin Lee; Da-Yeon Jeon; Su-Jin Sa; Yong-Dae Jeong; Ha-Seung Seong; Jung-Woo Choi; Shinichi Hochi; Eun-Seok Cho; Hak-Jae Chung

doi:10.5187/jast.2023.e27

Profiling of differentially expressed proteins between fresh and frozen-thawed Duroc boar semen using ProteinChip CM10

J Anim Sci Technol. 2023 Mar;65(2):401-411. doi: 10.5187/jast.2023.e27. Epub 2023 Mar 31.

Authors

Affiliations

¹ Planning and Coordination Division, National Institute of Animal Science, Wanju 55365, Korea.
² Swine Science Division, National Institute of Animal Science, Cheonan 31000, Korea.
³ TNT Research, Doosan Venture Digm, Anyang 14059, Korea.
⁴ Department of Animal Science, College of Animal Life Sciences, Kangwon National University, Chuncheon 24341, Korea.
⁵ Faculty of Textile Science and Technology, Shinshu University, Nagano 386-8567, Japan.

Abstract

Many studies have been conducted to improve technology for semen cryopreservation in pigs. However, computer-assisted analysis of sperm motility and morphology is insufficient to predict the molecular function of frozen-thawed semen. More accurate expression patterns of boar sperm proteins may be derived using the isobaric tags for relative and absolute quantification (iTRAQ) technique. In this study, the iTRAQ-labeling system was coupled with liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis to identify differentially expressed CM10-fractionated proteins between fresh and frozen-thawed boar semen. A total of 76 protein types were identified to be differentially expressed, among which 9 and 67 proteins showed higher and lower expression in frozen-thawed than in fresh sperm samples, respectively. The classified functions of these proteins included oxidative phosphorylation, mitochondrial inner membrane and matrix, and pyruvate metabolic processes, which are involved in adenosine triphosphate (ATP) synthesis; and sperm flagellum and motile cilium, which are involved in sperm tail structure. These results suggest a possible network of biomarkers associated with survival after the cryopreservation of Duroc boar semen.

Keywords: Boar semen; Isobaric tags for relative and absolute quantification (iTRAQ); ProteinChip; Semen cryopreservation; Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS).