Reyes A, Mercado E, Goicoechea B, Rosado A
Fertil Steril. 1976 Dec;27(12):1452-8. doi: 10.1016/s0015-0282(16)42263-6.
Changes in the concentration of -SH groups on the human and rabbit spermatozoal membrane and during epididymal maturation were studied by means of a new fluorescent probe, carboxyphenylmaleimide (CPhM), which reacts specifically with -SH groups. Binding of CPhM did not modify oxygen uptake, motility, or viability of the sperm cells used, but produced a characteristic increase in fluorescence. By analysis of this increase it was possible to calculate the presence of 35 +/- 4.2 and 55 +/- 8 nmoles of exposed -SH groups/10(8) rabbit and human ejaculated spermatozoa, respectively. Caput epididymal cells bound significantly more CPhM than did cauda epididymal cells or ejaculated spermatozoa (155 +/- 22, 78 +/- 11, and 35 +/- 4.2 nmoles/10(8) cells, respectively, in rabbit cells; and 184, 110 +/- 18, and 55 +/- 8 nmoles/10(8) cells, respectively, in humans cells). In addition to the differences in number of exposed -SH groups observed between human and rabbit sperm cells, the behavior of these membrane-reactive groups when ethylenediaminetetraacetate and/or zinc were added to the incubation media indicates that the participation of membrane--SH groups in sperm physiology is species-specific.
利用一种新型荧光探针羧基苯基马来酰亚胺(CPhM),研究了人及兔精子膜上-SH基团浓度在附睾成熟过程中的变化,该探针能与-SH基团特异性反应。CPhM的结合并不改变所用精子细胞的氧摄取、活力或生存能力,但会使荧光产生特征性增强。通过分析这种增强情况,分别计算出每10⁸个兔和人射出精子中暴露的-SH基团含量为35±4.2和55±8纳摩尔。附睾头细胞比附睾尾细胞或射出精子结合的CPhM显著更多(兔细胞中分别为155±22、78±11和35±4.2纳摩尔/10⁸个细胞;人细胞中分别为184、110±18和55±8纳摩尔/10⁸个细胞)。除了在人和兔精子细胞之间观察到的暴露-SH基团数量差异外,当向孵育培养基中添加乙二胺四乙酸和/或锌时,这些膜反应性基团的行为表明膜-SH基团在精子生理过程中的参与具有物种特异性。