Khumran A M, Yimer N, Rosnina Y, Wahid H, Ariff M O, Homayoun H, Asmatullah K, Bello T K
Department of Theriogenology and Production, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria, Nigeria.
Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
Vet World. 2019 Apr;13(4):649-654. doi: 10.14202/vetworld.2020.649-654. Epub 2020 Apr 11.
The aim of this study was to investigate the effects of different concentration of butylated hydroxytoluene (BHT) on sperm membrane surface protein "P25b" from cryopreserved bull semen in either lecithin based Bioxcell (BX) or two egg-yolk based extenders, tris-egg yolk (TEY), and citrate-egg yolk (CEY).
Forty-five semen samples, 15 each were extended with either BX, TEY, or CEY extender which contained different concentrations (0.0 - control, 0.5, 1.0, 1.5, 2.0, and 3.0 mM/mL) of BHT. The extended semen samples were frozen at a concentration of 20×10/mL in 0.25 mL straws and stored in liquid nitrogen for 2weeks. The frozen samples were thereafter thawed, proteins extracted and analyzed for quantities of protein P25b through direct sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel densitometry. Peptides were confirmed by Western blotting (WB).
Results showed that supplementation of BHT improved (p<0.05) quantity of protein P25b at concentrations of 0.5mM/mL for BX and at 1.0 mM/mL for TEY and CE when compared with the controls and other treatments.
BHT supplementation at 0.5 in BX and 1.0 mM/mL in TEY and CEY has protected bull sperm fertility marker protein P25b in frozen-thawed bull sperm.
本研究旨在调查不同浓度的丁基羟基甲苯（BHT）对基于卵磷脂的Bioxcell（BX）或两种基于蛋黄的稀释液（三羟甲基氨基甲烷 - 蛋黄（TEY）和柠檬酸盐 - 蛋黄（CEY））中冷冻保存的公牛精液精子膜表面蛋白“P25b”的影响。
45份精液样本，每份15份分别用含有不同浓度（0.0 - 对照、0.5、1.0、1.5、2.0和3.0 mM/mL）BHT的BX、TEY或CEY稀释液进行稀释。将稀释后的精液样本以20×10/mL的浓度冷冻在0.25 mL细管中，并在液氮中保存2周。此后，将冷冻样本解冻，提取蛋白质，并通过直接十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳凝胶密度测定法分析蛋白质P25b的含量。通过蛋白质印迹法（WB）确认肽段。
结果表明，与对照和其他处理相比，在BX中添加0.5 mM/mL的BHT以及在TEY和CE中添加1.0 mM/mL的BHT时，蛋白质P25b的含量有所提高（p<0.05）。
在BX中添加0.5 mM/mL以及在TEY和CEY中添加1.0 mM/mL的BHT可保护冻融公牛精子中的公牛精子生育力标记蛋白P25b。
