Parks J E, Hammerstedt R H
Biol Reprod. 1985 Apr;32(3):653-68. doi: 10.1095/biolreprod32.3.653.
Ram spermatozoa were obtained from different regions (caput, corpus, and cauda) of the epididymis and their plasma membrane was removed using a nitrogen cavitation treatment (750 psi, 10 min equilibration at 4 degrees C). Membrane was recovered after sucrose gradient centrifugation and identified using 125I-succinylated concanavalin A (125I-succConA) as a surface marker. Based on fluorescein isothiocyanate-succConA (FITC-succConA) labeling and electron microscopy, cavitation removed plasma membrane from the anterior sperm head in the area overlying the acrosome. Cholesterol was the major sterol in plasma membrane, with desmosterol present in sperm entering the epididymis (caput sperm) but negligible in sperm after epididymal transit (cauda sperm). Ethanolamine and choline phosphoglycerides represented 70-80% of membrane phospholipids, with the ethanolamine fraction decreasing relative to choline phosphoglycerides during epididymal transit. The molar ratio of cholesterol to phospholipid increased in the plasma membrane during maturation. The bulk phospholipid-bound fatty acids consisted primarily of palmitoyl acyl groups (16:0) in caput sperm and docosahexaenoyl acyl groups (22:6) in cauda sperm. The choline phosphoglyceride fraction was purified and analyzed. It consisted of a mixture of ether acyl glycero-3-phosphocholine and diacyl phosphoglyceride, with the dominant acyl residue, at all stages of epididymal maturation, being 22:6 throughout epididymal transit. The significance of these findings relative to acquisition of fertilization capacity by sperm during epididymal maturation is discussed.
从附睾的不同区域(头部、体部和尾部)获取公羊精子,并用氮气空化处理(750磅力/平方英寸,4℃平衡10分钟)去除其质膜。经蔗糖梯度离心后回收膜,并使用125I-琥珀酰化伴刀豆球蛋白A(125I-succConA)作为表面标记物进行鉴定。基于异硫氰酸荧光素-succConA(FITC-succConA)标记和电子显微镜观察,空化去除了顶体上方区域精子头部前端的质膜。胆固醇是质膜中的主要固醇,在进入附睾的精子(头部精子)中存在去氢胆固醇,但在附睾转运后的精子(尾部精子)中可忽略不计。乙醇胺和胆碱磷酸甘油酯占膜磷脂的70-80%,在附睾转运过程中,乙醇胺部分相对于胆碱磷酸甘油酯减少。在成熟过程中,质膜中胆固醇与磷脂的摩尔比增加。大量与磷脂结合的脂肪酸在头部精子中主要由棕榈酰基(16:0)组成,在尾部精子中主要由二十二碳六烯酰基(22:6)组成。对胆碱磷酸甘油酯部分进行了纯化和分析。它由醚酰甘油-3-磷酸胆碱和二酰基磷酸甘油酯的混合物组成,在附睾成熟的所有阶段,主要的酰基残基在整个附睾转运过程中均为22:6。讨论了这些发现相对于精子在附睾成熟过程中获得受精能力的意义。