Srivastava A, Olson G E
Department of Cell Biology, Vanderbilt University, Nashville, TN 37232.
Mol Reprod Dev. 1991 Aug;29(4):357-64. doi: 10.1002/mrd.1080290407.
To investigate surface glycoprotein changes during post-testicular maturation, plasma membranes were isolated from proximal caput, distal caput, and cauda epididymal rat spermatozoa. Membrane glycoproteins were identified on Western blots of SDS-PAGE fractionated samples using biotinylated lectins and Vecta-stain reagents; these were compared to glycoproteins present in cauda epididymal luminal fluid. Lens culinaris agglutinin, Pisum sativum agglutinin, peanut agglutinin, wheat germ agglutinin, Ricinus communis agglutinin, Ulaex europaeus agglutinin, and Dolichol biflorus agglutinin each bound a specific subset of the polypeptides present. Several types of glycoprotein changes were noted including their appearance, loss, alteration of staining intensity, and alteration of electrophoretic mobility. Some maturation-dependent sperm surface glycoproteins co-migrated with glycoproteins present in epididymal fluid. This approach of direct analysis of the glycoproteins in purified plasma membranes identifies a broader spectrum of maturation-related surface changes occurring within the epididymis than are noted with surface labeling procedures.
为了研究睾丸后成熟过程中表面糖蛋白的变化,从大鼠附睾头近端、附睾头远端和附睾尾的精子中分离出质膜。使用生物素化凝集素和Vecta染色试剂,在SDS-PAGE分级分离样品的Western印迹上鉴定膜糖蛋白;将这些糖蛋白与附睾尾管腔液中存在的糖蛋白进行比较。刀豆凝集素、豌豆凝集素、花生凝集素、麦胚凝集素、蓖麻凝集素、欧洲刺柏凝集素和双花扁豆凝集素分别结合了存在的特定多肽亚群。观察到几种类型的糖蛋白变化,包括它们的出现、消失、染色强度的改变和电泳迁移率的改变。一些成熟依赖性精子表面糖蛋白与附睾液中存在的糖蛋白共同迁移。这种直接分析纯化质膜中糖蛋白的方法,比表面标记程序能鉴定出附睾内发生的更广泛的与成熟相关的表面变化。