THE EFFECT OF SULFHYDRYL OXIDATION ON THE MORPHOLOGY OF IMMATURE HAMSTER EPIDIDYMAL SPERMATOZOA INDUCED TO ACQUIRE MOTILITY INVITRO
THE EFFECT OF SULFHYDRYL OXIDATION ON THE MORPHOLOGY OF IMMATURE HAMSTER EPIDIDYMAL SPERMATOZOA INDUCED TO ACQUIRE MOTILITY INVITRO
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DOI：
10.1095/biolreprod39.1.141
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发表时间：
1988-08-01
影响因子：
3.6
通讯作者：
CHANG, TSK
中科院分区：
文献类型：
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作者：
CORNWALL, GA;VINDIVICH, D;CHANG, TSK
附睾头的静止精子在含有茶碱、精浆和白蛋白的培养液中孵育后逐渐活动。我们以前曾报道，在这些培养条件下，诱导获得活力的精子表现出明显的鞭毛角，精子头部或中段弯曲90-180度。朝向尾巴。此外，我们还证明了在运动诱导介质中加入巯基氧化剂联胺时，精子鞭毛不会发生弯曲。在本研究中，我们进一步研究了在体外诱导获得运动能力的未成熟头部精子的形态和巯基含量的变化。我们发现联胺以剂量依赖的方式阻止鞭毛弯曲和维持精子鞭毛的直线度。此外，使用巯基试剂单溴双胺(MBBR)对精子头部的巯基进行的流式细胞术分析表明，1)联胺氧化头部精子的巯基，以及2)在能够防止精子成角的联胺浓度下，只有不到15%的反应精子的巯基被氧化。另一种巯基氧化剂四硫酸钠(Natt)和含有巯基氧化酶活性的仓鼠附睾液(CEF)在诱导的头部精子和氧化精子中也保持了鞭毛的直线度。然而，用巯基氧化剂处理的头部精子的鞭毛是暂时的；随着孵化时间的延长，联胺或CEF处理的精子表现出鞭毛弯曲。进一步的研究表明，在诱导介质中加入谷胱甘肽还原酶抑制剂呋喃妥因时，观察到的巯基氧化精子的鞭毛直线度是持续的。对呋喃妥因处理的精子进行的流式细胞仪分析表明，呋喃妥因维持了由联胺形成的精子二硫化物，并阻止了精子二硫化物还原为巯基。综上所述，这些研究表明，在体外诱导获得运动能力的未成熟附睾头精子，在维持其形态方面具有重要意义，并提示在体内精子成熟过程中，巯基氧化可能在精子运动能力的发育过程中起重要作用。
Immotile spermatozoa from the caput epididymidis become progressively motile when incubated in medium containing theophylline, seminal plasma, and albumin. We previously reported that under these incubation conditions the spermatozoa induced to acquire motility exhibited a marked flagellar angularity, with the sperm head or midpiece bent 90-180.degree. towards the tail. In addition, we demonstrated that sperm flagellar bending did not occur when the sulfhydryl oxidant diamide was added to the motility induction medium. In the present study, we examined further the effect of sulfhydryl oxidation on the morphology and sulfhydryl content of immature caput spermatozoa induced to acquire motility in vitro. We found that flagellar bending was prevented and sperm flagellar straightness was maintained in dose-dependent manner by diamide. Moreover, flow cytometric analysis of caput sperm sulfhydryls using the sulfhydryl reagent monobromobimane (mBBr) revealed that 1) diamide oxidizes caput sperm sulfhydryls, and 2) less than 15% of the total reactive sperm sulfhydryls were oxidized at diamide concentrations capable of preventing sperm angulation. Sodium tetrathionate (NaTT), another sulfhdryl oxidant, and hamster cauda epididymal fluid (CEF) containing sulfhydryl oxidase enzyme activity also maintained flagellar straightness in induced caput spermatozoa and oxidized sperm sulfhydryls. The flagellar straightness in caput spermatozoa treated with sulfhydryl oxidants, however, was temporary; with extended incubation, diamide- or CEF-treated spermatozoa exhibited flagellar bending. Additional studies showed that the flagellar straightness observed in sulfhydryl-oxidized spermatozoa was sustained when nitrofurantoin, an inhibitor of glutathione reductase, was included in the induction medium. Flow cytometric analysis of nitrofurantoin-treated spermatozoa showed that nitrofurantoin maintained the sperm disulfides formed by diamide and prevented the reduction of sperm disulfides back to sulfhydryls. Taken together, these studies demonstrate the significance of sulfhydryl oxidation in maintaining the morphology of immature caput epididymal spermatozoa induced to acquire motility in vitro and suggest that sulfhydryl oxidation may be important in the development of motility during sperm epididymal maturation in vivo.