Metabolomic analysis of seminal plasma to identify goat semen freezability markers

Bingbing Xu; Xue Bai; Jian Zhang; Boyuan Li; Yanjun Zhang; Rui Su; Ruijun Wang; Zhiying Wang; Qi Lv; Jiaxin Zhang; Jinquan Li

doi:10.3389/fvets.2023.1132373

Metabolomic analysis of seminal plasma to identify goat semen freezability markers

Front Vet Sci. 2023 Mar 10:10:1132373. doi: 10.3389/fvets.2023.1132373. eCollection 2023.

Authors

Bingbing Xu¹, Xue Bai¹, Jian Zhang¹, Boyuan Li¹, Yanjun Zhang¹, Rui Su¹, Ruijun Wang¹, Zhiying Wang¹, Qi Lv¹, Jiaxin Zhang^{1

2

3}, Jinquan Li^{1

2

3

4}

Affiliations

¹ College of Animal Science, Inner Mongolia Agricultural University, Hohhot, China.
² Key Laboratory of Animal Genetics, Breeding and Reproduction, Hohhot, China.
³ Key Laboratory of Mutton Sheep Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Hohhot, China.
⁴ Inner Mongolia Jinlai Animal Husbandry Technology Co., Ltd., Hohhot, China.

Abstract

Factors affecting sperm freezability in goat seminal plasma were investigated. Based on the total motility of thawed sperm, goats were divided into a high-freezability (HF) group with >60% total motility (n = 8) and a low-freezability (LF) group with <45% total motility (n = 8). Sperm and seminal plasma from the HF and LF groups were separated, HF seminal plasma was mixed with LF spermatozoa, LF seminal plasma was mixed with HF sperm, and the products were subjected to a freeze-thaw procedure. Semen from individual goats exhibited differences in freezability. HF semen had higher sperm motility parameters and plasma membrane and acrosome integrity after thawing; this difference could be related to the composition of seminal plasma. Seminal plasma from the HF and LF groups was evaluated using metabolomic analysis, and multivariate statistical analysis revealed a clear separation of metabolic patterns in the seminal plasma of goats with different freezability classifications. Forty-one differential metabolites were identified using the following screening conditions: variable importance in the projection > 1 and 0.05 < P-value < 0.1. Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed significant enrichment of central carbon metabolism in cancer, protein digestion and absorption, aminoacyl-tRNA, and other pathways and significant differences in the abundance of seven differential metabolites, including L-glutamine, L-aspartate, L-arginine, phenylpyruvate, benzoic acid, ketoisocaproic acid, and choline between seminal plasma from the HF and LF groups (P-value < 0.05). These significantly differentially-expressed metabolites may be potential biomarkers for sperm freezability. L-glutamine, L-aspartate, and L-arginine may directly affect sperm freezability. Benzoic acid, ketoisocaproic acid, and choline may regulate sperm freezability by participating in anabolic processes involving phenylalanine, leucine, and phosphatidylcholine in sperm.

Keywords: cryopreservation; freezability; goat spermatozoa; metabolomic; seminal plasma.

Grants and funding

This research was funded by the National Key R&D Program of China, Grant No. 2021YFD1300901, the China Agriculture Research System of MOF and MARA (No. CARS-39-06), and Program for Innovative Research Team in Universities of Inner Mongolia Autonomous Region (NMGIRT2322).