Skudlarek M D, Orgebin-Crist M C
Center for Reproductive Biology Research, Vanderbilt School of Medicine, Nashville, Tennessee 37232.
J Reprod Fertil. 1988 Nov;84(2):611-7. doi: 10.1530/jrf.0.0840611.
Each epididymis of control and swainsonine-fed rats (5 micrograms/ml drinking water) was divided into 5 segments, and tissue, spermatozoa and sperm-free supernatants were prepared from each segment. When levels of 3 lysosomal glycosidases and total protein were determined, the proximal cauda contained the greatest concentration of glycosidase. The specific-activity profile for beta-glucuronidase and beta-galactosidase was similar in swainsonine-fed and control rats. However, the concentration of alpha-D-mannosidase in tissue of all segments was significantly greater in swainsonine-fed rats than in age-matched controls. Enzyme activity for alpha-D-mannosidase after swainsonine treatment was significantly greater in spermatozoa from the caput, than in spermatozoa from the corpus and the cauda epididymidis. Since the alpha-D-mannosidase activity was optimal at pH 4.5 and studies with highly specific antibody to lysosomal alpha-D-mannosidase immunoprecipitated all of the alpha-D-mannosidase present in detergent extracts of epididymal tissue, spermatozoa, and sperm-free supernatant, the enzyme studied is of lysosomal origin.
将对照大鼠和喂食苦马豆素的大鼠（饮用水中含5微克/毫升）的每个附睾分为5段，并从每段制备组织、精子和无精子上清液。测定3种溶酶体糖苷酶和总蛋白水平时，附睾近尾段的糖苷酶浓度最高。喂食苦马豆素的大鼠和对照大鼠中β-葡萄糖醛酸酶和β-半乳糖苷酶的比活性谱相似。然而，喂食苦马豆素的大鼠所有段组织中的α-D-甘露糖苷酶浓度显著高于年龄匹配的对照大鼠。苦马豆素处理后，附睾头精子中的α-D-甘露糖苷酶活性显著高于附睾体和附睾尾精子中的活性。由于α-D-甘露糖苷酶活性在pH 4.5时最佳，且用针对溶酶体α-D-甘露糖苷酶的高度特异性抗体进行的研究免疫沉淀了附睾组织、精子和无精子上清液去污剂提取物中存在的所有α-D-甘露糖苷酶，因此所研究的酶起源于溶酶体。
