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Abstract：
In the present study, there was evaluation of cryocapacitation-associated changes, apoptotic-like changes, deprotamination, total antioxidant capacity (TAC), and in vitro sperm functional attributes in Barbari bucks after freezing-thawing. The correlation between deprotamination and sperm functional characteristics was established. Using immunoblotting procedures, there was detection of the presence of a single 28-kDa protein band corresponding to protamine-1. The localization in the head region of the spermatozoa was further validated by an immunofluorescence test. Capacitated (B-) and acrosome-reacted (AR-) pattern spermatozoa, spermatozoa with the externalization of phosphatidylserine and a relatively lesser mitochondrial transmembrane potential, and deprotamination and DNA fragmentation was greater (P < 0.05) after freezing-thawing and indicated there were cryocapacitation- and apoptotic-like changes, respectively. Furthermore, the detection of phosphorylation of tyrosine-containing proteins with use of immunoblotting and immunofluorescence procedures confirmed there were cryocapacitation-like changes in the buck spermatozoa after freezing-thawing. Total antioxidant capacity (TAC), in vitro thermal resistance response, Vanguard distance, progesterone sensitivity, and in vitro capacitation response were less (P < 0.05) in the spermatozoa after freezing-thawing compared with spermatozoa after initial dilution and equilibration. Deprotamination (chromomycin A3-positive cells, CMA3+) and DNA fragmentation (TUNEL+ve) were positively correlated with B- and AR-pattern spermatozoa, while other values for other variables were negatively correlated. In conclusion, the results of this study indicated there was protamine-1 in buck spermatozoa and after freezing-thawing there was a loss of protamine-1 combined with cryocapacitation-associated changes and apoptotic-like changes in buck spermatozoa. Spermatozoa deprotamination might be attributed to increased DNA fragmentation, resulting in compromised fertilizing capacity of buck spermatozoa.
摘要：
在本研究中,评估了与冷冻电容相关的变化，凋亡样变化，去质子化，总抗氧化能力(TAC)，冻融后Barbari雄鹿的体外精子功能属性。建立了去质子化与精子功能特征之间的相关性。使用免疫印迹程序，检测到对应于鱼精蛋白-1的单个28-kDa蛋白带的存在。通过免疫荧光测试进一步验证了精子头部区域的定位。电容（B-）和顶体反应（AR-）模式精子，精子具有磷脂酰丝氨酸的外化和相对较小的线粒体跨膜电位，冻融后去质子化和DNA片段化较大（P<0.05），表明有冷冻和凋亡样变化，分别。此外，使用免疫印迹和免疫荧光程序检测含酪氨酸蛋白的磷酸化，证实了冷冻-解冻后buck精子中存在类似冷冻电容的变化。总抗氧化能力（TAC），体外热阻反应，前进距离,孕酮敏感性,与初始稀释和平衡后的精子相比，冻融后精子的体外获能反应较低（P<0.05）。去质子化（色霉素A3阳性细胞，CMA3+)和DNA片段化(TUNEL+ve)与B-和AR-型精子呈正相关,而其他变量的其他值呈负相关。总之,这项研究的结果表明，在buck精子中存在鱼精蛋白-1，冻融后，鱼精蛋白-1的丢失，并伴有与冷冻电容相关的变化和凋亡样变化。精子去质子化可能归因于DNA片段化增加，导致降压精子的受精能力受损。
