Oberländer G, Yeung C H, Cooper T G
Institute of Reproductive Medicine, M-unster, Germany.
J Reprod Fertil. 1996 Mar;106(2):231-9. doi: 10.1530/jrf.0.1060231.
The chlortetracycline fluorescence assay was used to study the status of capacitation and the extent of induced acrosome reactions in cauda epididymidal spermatozoa from fertile and infertile rats fed, respectively, with vehicle or ornidazole (400 mg kg-1 day-1) for 10 days. Uniform bright fluorescence over the whole head was classified as the uncapacitated pattern, whereas a postacrosomal dark band, and a uniformly weaker fluorescence over the acrosome, reflected patterns intermediate between the uncapacitated and acrosome-reacted states. Acrosome-reacted spermatozoa displayed a dark head but always retained fluorescence at their tip. There was no difference between experimental and control groups of rats with regard to the development of the chlortetracycline fluorescence patterns during incubation. Under basal incubation conditions, the acrosome reaction was slightly delayed in spermatozoa from ornidazole-treated animals. In contrast, more spermatozoa were acrosome reacted in this group after incubation for 5 h when the concentration of BSA was increased from 4 to 20 mg ml-1. The Ca(2+)-ionophore A23187 induced a similar stimulation of capacitation and acrosome reactions in spermatozoa from control and ornidazole-fed animals, but in the latter group A23187 caused strong immobilization of spermatozoa. In the capacitation medium containing 5 mmol lactate l-1 and 5 mmol glucose l-1, the straight-line velocity of spermatozoa from ornidazole-treated rats was reduced by 50% compared with controls, irrespective of the concentration of BSA. Two glycolytic enzymes, triose phosphate isomerase and glyceraldehyde 3-phosphate dehydrogenase, displayed reduced activity (48% and 68% of controls, respectively) in cauda epididymidal spermatozoa from ornidazole-fed rats, whereas the activities of hexokinase and lactate dehydrogenase remained unchanged. This finding suggests that the fertility-compromising action of ornidazole is due to a disturbed glycolytic pathway.
采用金霉素荧光分析法，研究分别用赋形剂或奥硝唑（400毫克/千克/天）喂养10天的可育和不育大鼠附睾尾精子的获能状态及诱导顶体反应的程度。整个头部均匀明亮荧光被归类为未获能模式，而顶体后暗带以及顶体上均匀较弱的荧光反映了介于未获能和顶体反应状态之间的模式。顶体反应的精子头部呈暗色，但顶端始终保留荧光。在孵育过程中，实验组和对照组大鼠的金霉素荧光模式发展没有差异。在基础孵育条件下，奥硝唑处理动物的精子顶体反应略有延迟。相比之下，当牛血清白蛋白浓度从4毫克/毫升增加到20毫克/毫升时，该组孵育5小时后更多精子发生了顶体反应。钙离子载体A23187在对照组和奥硝唑喂养动物的精子中诱导了类似的获能和顶体反应刺激，但在后者组中，A23187导致精子强烈制动。在含有5毫摩尔/升乳酸和5毫摩尔/升葡萄糖的获能培养基中，无论牛血清白蛋白浓度如何，奥硝唑处理大鼠的精子直线速度比对照组降低了50%。两种糖酵解酶，磷酸丙糖异构酶和3 - 磷酸甘油醛脱氢酶，在奥硝唑喂养大鼠的附睾尾精子中活性降低（分别为对照组的48%和68%），而己糖激酶和乳酸脱氢酶的活性保持不变。这一发现表明，奥硝唑损害生育能力的作用是由于糖酵解途径紊乱。
