Dept. of Theriogenology, Faculty of Veterinary Medicine, Benha University, Egypt.
Department of Veterinary Science, University of Torino, Grugliasco, Italy.
Theriogenology. 2022 Oct 15;192:141-149. doi: 10.1016/j.theriogenology.2022.08.022. Epub 2022 Aug 23.
This work aimed to determine the effect of cysteamine (25, 50, 100 and 200 μM) incorporated during dilution on frozen thawed buffalo semen quality. Semen was collected twice weekly for 7 consecutive weeks from three Egyptian buffalo bulls using an artificial vagina. Semen samples were pooled and extended with a Tris-based extender, cooled, equilibrated and finally frozen in liquid nitrogen. The diluted semen was evaluated for motility, viability, morphology, plasma membrane and DNA integrity, in addition to oxidative stress and in vitro fertilizing capability. The post thaw motility and velocity parameters noticeably increased with different concentrations of cysteamine (mainly 100 μM) during different incubation periods. The post thaw sperm viability and normality significantly (p < 0.05) improved with concentrations of 50 and 100 μM. Plasma membrane integrity substantially increased at all concentrations of cysteamine. Cysteamine reduced alanine aminotransferase (at all concentrations), aspartate aminotransferase (at 25-100 μM), and creatine kinase (at 100 and 200 μM). Cysteamine at a concentration of 100 μM noticeably enhanced the total antioxidant capacity and glutathione peroxidase and decreased nitric oxide production. Cysteamine, at concentrations of 100 and 200 μM, increased the DNA intensity in the comet head (%) and decreased the DNA % in the comet tail. The comet tail length and moment substantially decreased at concentrations of 50-200 μM. Cysteamine did not affect the in vitro fertilizing capability of sperm. In conclusion, cysteamine incorporation (mainly at a concentration of 100 μM) in buffalo semen extender showed varying protective effects on different sperm parameters against cryo-damage; however, it did not affect the in vitro fertilizing capacity of sperm.
本研究旨在确定半胱胺(25、50、100 和 200 μM)在稀释过程中对冷冻解冻水牛精液质量的影响。使用人工阴道每周从三头埃及公牛中采集两次精液,连续采集 7 周。将精液样本混合并使用基于 Tris 的稀释液进行扩展,然后冷却、平衡,最后在液氮中冷冻。评估稀释精液的活力、活力、形态、质膜和 DNA 完整性,以及氧化应激和体外受精能力。不同浓度的半胱胺(主要是 100 μM)在不同孵育期显著增加了解冻后的运动和速度参数。浓度为 50 和 100 μM 的半胱胺显著提高了解冻后精子的活力和正常率。质膜完整性在所有半胱胺浓度下都显著增加。半胱胺降低了丙氨酸氨基转移酶(所有浓度)、天冬氨酸氨基转移酶(25-100 μM)和肌酸激酶(100 和 200 μM)。浓度为 100 μM 的半胱胺显著增强了总抗氧化能力和谷胱甘肽过氧化物酶,降低了一氧化氮的产生。浓度为 100 和 200 μM 的半胱胺增加了彗星头中的 DNA 强度(%),并降低了彗星尾中的 DNA%。在 50-200 μM 浓度下,彗星尾长度和矩显著降低。半胱胺对精子的体外受精能力没有影响。总之,半胱胺(主要在 100 μM 浓度下)在水牛精液稀释液中的加入对不同的精子参数对冷冻损伤的保护作用不同;然而,它并没有影响精子的体外受精能力。