Tramer F, Rocco F, Micali F, Sandri G, Panfili E
Department of Biochemistry, Biophysics and Macromolecular Chemistry, University of Trieste, Trieste, Italy.
Biol Reprod. 1998 Oct;59(4):753-8. doi: 10.1095/biolreprod59.4.753.
Mammalian caput and cauda epididymidal spermatozoa exhibit diverse stages of maturation, and their plasma membrane shows diverse composition and stability levels, thus enabling these spermatozoa to undergo the acrosomal reaction after transit through the epididymis. As a result, the study of antiperoxidative mechanisms is quite relevant, since epididymal spermatozoa must be properly protected against agents such as reactive oxygen species, which can impair the complex maturation process. We considered activities of certain enzymes (glutathione peroxidase [GPx], phospholipid hydroperoxide glutathione peroxidase [PHGPx], glutathione reductase [GR], superoxide dismutase [SOD], and catalase [CAT]) and the vitamin E content in isolated rat caput and cauda epididymidal spermatozoa. The results indicate that caput epididymidal sperm have significantly greater PHGPx (3.5x), GPx (2.4x), and SOD (1.7x) activities, as well as a greater amount of vitamin E (3.8x). There were no detectable differences in the GR and CAT activities of caput and cauda epididymidal spermatozoa. The substantial drop in PHGPx activity during epididymal transit is discussed in relation to an additional function of this enzyme: the use of caput sperm protamines as a sulfhydryl substrate. In vitro peroxidation of the two sperm populations by the free radical generator (azo-initiator) 2,2'-azobis(2-amidinopropane) dihydrochloride revealed that only about 13% of the vitamin E content of the caput epididymidal spermatozoa was consumed, which contrasts with the greater consumption (about 70%) of the vitamin in cauda epididymidal spermatozoa. Selective inhibition of PHGPx, SOD, or CAT did not change this picture. The higher susceptibility of cauda epididymidal spermatozoa to radicals is discussed in relation to the diverse enzymatic activities, vitamin E content, and peroxidative response. These factors are correlated with the different stages of sperm cell maturation, which are characterized-from caput to cauda epididymidis-by progressive destabilization of the plasma and acrosomal membranes.
哺乳动物附睾头和附睾尾精子呈现出不同的成熟阶段,其质膜具有不同的组成和稳定性水平,从而使这些精子在通过附睾后能够发生顶体反应。因此,抗过氧化机制的研究具有重要意义,因为附睾精子必须得到妥善保护,免受活性氧等物质的侵害,这些物质会损害复杂的成熟过程。我们研究了分离出的大鼠附睾头和附睾尾精子中某些酶(谷胱甘肽过氧化物酶[GPx]、磷脂氢过氧化物谷胱甘肽过氧化物酶[PHGPx]、谷胱甘肽还原酶[GR]、超氧化物歧化酶[SOD]和过氧化氢酶[CAT])的活性以及维生素E的含量。结果表明,附睾头精子的PHGPx(3.5倍)、GPx(2.4倍)和SOD(1.7倍)活性显著更高,维生素E含量也更高(3.8倍)。附睾头和附睾尾精子的GR和CAT活性没有可检测到的差异。文中讨论了附睾转运过程中PHGPx活性的大幅下降与该酶的另一个功能有关:将附睾头精子的鱼精蛋白用作巯基底物。自由基发生器(偶氮引发剂)2,2'-偶氮双(2-脒基丙烷)二盐酸盐对这两种精子群体进行的体外过氧化反应显示,附睾头精子中只有约13%的维生素E含量被消耗,这与附睾尾精子中维生素E的更大消耗量(约70%)形成对比。对PHGPx、SOD或CAT的选择性抑制并没有改变这种情况。文中讨论了附睾尾精子对自由基更高的敏感性与不同的酶活性、维生素E含量和过氧化反应有关。这些因素与精子细胞成熟的不同阶段相关,从附睾头到附睾尾,其特征是质膜和顶体膜逐渐不稳定。