Isolation of a stable apical segment of the guinea pig sperm acrosome.
Isolation of a stable apical segment of the guinea pig sperm acrosome.
复制标题
豚鼠精子顶体稳定顶端段的分离。
DOI：
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
R. Hyne
中科院分区：
文献类型：
--
作者：
A. Stojanoff;H. Bourne;A. Andrews;R. Hyne
机械分离豚鼠精子顶体的大顶端段，然后用密度梯度离心分离。当分离的顶体帽悬浮在pH为5.6的蔗糖基缓冲培养基中，无论是否添加顶蛋白抑制剂对氨基苯脒（pAB），顶体帽都非常稳定，保持新月形形态。电镜检查显示顶体帽无质膜，由一不连续的顶体外膜结合。酶解后的酶分析表明，顶胞苷和透明质酸酶具有较高的比活性，而酸性磷酸酶只有微量活性，没有芳基硫酸酯酶、磷脂酶A2和磷脂酶C活性。如果在没有pAB的情况下分离后立即检测，可以在分离的顶体帽中检测到显著的颗粒顶蛋白活性，但只有微量的可溶性顶蛋白活性。用10%甘油在低pH （pH 3.0）下缓冲提取顶体帽后，获得了高比活性的可溶性顶蛋白活性。新方法提供了一种快速分离纯化豚鼠精子顶体片段的方法。
The large apical segments of guinea pig sperm acrosomes were mechanically separated from the spermatozoa and subsequently isolated by density gradient centrifugation. The isolated acrosomal caps were very stable and maintained their crescent morphology when suspended in sucrose-based medium buffered at pH 5.6, with or without the acrosin inhibitor p-aminobenzamidine (pAB). Examination under the electron microscope showed that the acrosomal caps were free of plasma membrane and were bound by an outer acrosomal membrane which was discontinuous. Enzymatic analysis after lysis of the caps indicated that acrosin and hyaluronidase were present with high specific activity, while only a trace amount of acid phosphatase activity and no arylsulphatase, phospholipase A2, or phospholipase C activities were present. Significant particulate acrosin activity, but only trace amounts of soluble acrosin activity, could be detected in the isolated acrosomal caps if assayed immediately after isolation in the absence of pAB. However, soluble acrosin activity of high specific activity was obtained after the acrosomal caps were extracted by 10% glycerol buffered at low pH (pH 3.0). The new procedures provide a means to isolate and purify guinea pig sperm apical acrosomal segments rapidly.
与牛精子顶体外膜相关的大分子复合物的分离和表征。
影响因子：
3.6
作者：
Olson,GE;Winfrey,VP;Garbers,DL;Noland,TD
通讯作者：
Noland,TD
豚鼠精子在pH 7.8以下缓冲介质中获能和顶体反应所需的血清因子。
影响因子：
3.6
作者：
Hyne,RV;Garbers,DL
通讯作者：
Garbers,DL
兔精子顶体的碱性磷酸酶和磷脂酶 C 水解对硝基苯磷酰胆碱。
DOI：
10.1016/0006-291x(82)92116-7
发表时间：
1982
影响因子：
3.1
作者：
Srivastava,PN;Brewer,JM;WhiteJr,RA
通讯作者：
WhiteJr,RA