Development changes occurring in the lipids of ram epididymal spermatozoa plasma membrane.
Development changes occurring in the lipids of ram epididymal spermatozoa plasma membrane.
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DOI：
10.1095/biolreprod32.3.653
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发表时间：
1985-04
影响因子：
3.6
通讯作者：
J. Parks;R. Hammerstedt
中科院分区：
文献类型：
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作者：
J. Parks;R. Hammerstedt
从附睾的不同区域（头、体和尾）获得公羊精子，并使用氮空化处理（750 psi，4°C 平衡 10 分钟）去除其质膜。蔗糖梯度离心后回收膜，并使用 125I-琥珀酰化刀豆球蛋白 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，空化作用从顶体上方区域的前精子头部去除了质膜。胆固醇是质膜中的主要甾醇，进入附睾的精子（带头精子）中存在去氨甾醇，但在附睾转运后的精子（尾精子）中含量可忽略不计。乙醇胺和磷酸胆碱甘油酯占膜磷脂的 70-80%，在附睾转运过程中乙醇胺比例相对于磷酸胆碱甘油酯有所减少。在成熟过程中，质膜中胆固醇与磷脂的摩尔比增加。大量磷脂结合脂肪酸主要由头精子中的棕榈酰酰基（16:0）和尾精子中的二十二碳六烯酰酰基（22:6）组成。纯化并分析磷酸甘油胆碱酯级分。它由醚酰基甘油-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，在整个附睾运输过程中，酰基残基占主导地位，比例为22:6。讨论了这些发现相对于附睾成熟过程中精子获得受精能力的重要性。
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.