海藻糖替代部分甘油对延边黄牛冻融精子质量的影响

doi:10.16431/j.cnki.1671-7236.2022.01.028

Abstract

Abstract: [Objective] This study was aimed to explore whether the concentration of glycerol in cryoprotectant could be reduced by supplementing trehalose, so as to improve the quality of frozen-thawed sperm. [Method] Sperm were treated with 6% glycerol (group Ⅰ) and 3% glycerol+0, 50, 100 and 150 mmol/L trehalose (Ⅱ, Ⅲ, Ⅳ and Ⅴ groups), respectively. The motility, plasma membrane integrity, acrosome integrity and dynamic parameters of frozen-thawed sperms were detected by the dynamic (static) image detection system (CASA), and the best trehalose treatment concentration was selected for subsequent experiments. Tyrosine phosphorylation of sperm protein was detected by Western blotting, the viability and mitochondrial membrane potential of sperm were detected by Hoechst 33342/PI/JC-1 combined staining, sperm DNA integrity was detected by chromomycin A₃ (CMA₃) staining, and the level of lipid disorder in sperm membrane was detected by cyanine 540 (M540) and Yo-Pro-1 staining. [Result] Compared with Ⅰ group, sperm motility, plasma membrane and acrosomal membrane integrity, VCL (curvilinear velocity) and STR (straightness) of Ⅱ group were significantly decreased (P<0.05), the sperm motility, VCL, VSL (straight line velocity), VAP (average path velocity), LIN (linearity) and STR of Ⅲ, Ⅳ and Ⅴ groups were significantly increased (P<0.05). Therefore, 100 mmol/L trehalose was used to treat sperm in the follow-up test. Compared with fresh sperm, the protein tyrosine phosphorylation bands of frozen-thawed sperm was decreased significantly after capacitation (P<0.05), and the viability and mitochondrial membrane potential of frozen-thawed sperm was decreased significantly (P<0.05). The level of high membrane lipid disorder of frozen-thawed live sperm and dead sperm was increased significantly (P<0.05), and 100 mmol/L trehalose could significantly reduce the level of high membrane lipid disorder (P<0.05). The protamine deletion rate of frozen-thawed sperm was significantly increased (P<0.05), and the protamine deletion rate of Ⅱ and 100 mmol/L trehalose groups was significantly lower than that of Ⅰ group (P<0.05). [Conclusion] Trehalose could be used as a new cryoprotectant to reduce the toxicity of glycerol. Replacing part of glycerol with 100 mmol/L trehalose could significantly improve the sperm quality of Yanbian Yellow cattle after freezing and thawing.

Key words: Yanbian Yellow cattle; frozen-thawed; trehalose; glycerol; sperm quality

CLC Number:

S823.8

CHEN Xuan, LYU Yanqiu, HAN Yue, XU Yichao, JIN Yi. Effect of Trehalose Replacing Partial Glycerol on Frozen-thawed Sperm Quality of Yanbian Yellow Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 256-264.

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Effect of Trehalose Replacing Partial Glycerol on Frozen-thawed Sperm Quality of Yanbian Yellow Cattle

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