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首页> 外文期刊>Biochemistry >DISTRIBUTION AND DYNAMICS OF MOUSE SPERM SURFACE GALACTOSYLTRANSFERASE - IMPLICATIONS FOR MAMMALIAN FERTILIZATION
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DISTRIBUTION AND DYNAMICS OF MOUSE SPERM SURFACE GALACTOSYLTRANSFERASE - IMPLICATIONS FOR MAMMALIAN FERTILIZATION

机译：小鼠精子表面半乳糖基转移酶的分布和动力学-对哺乳动物的受精意义

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It has been proposed that a mouse sperm surface beta-1,4-galactosyltransferase functions as a receptor for the zona pellucida during fertilization. In this paper we used two monovalent fluorescent probes specific for galactosyltransferase: a trinitrophenylated derivative of UDP-galactose and rhodaminated alpha-lactalbumin. We found that galactosyltransferase was initially present over the posterior head of acrosome-intact sperm but became progressively localized to the plasma membrane overlying the acrosomal region after it was cross-linked with an anti-galactosyltransferase polyclonal antibody. Labeled mouse sperm that were treated with the calcium ionophore A23187 revealed that galactosyltransferase remained on the posterior head after acrosomal exocytosis. However, if galactosyltransferase was first cross-linked and redistributed with antibody and then acrosome reacted with A23187, all head fluorescence was lost. In addition, although anti-galactosyltransferase antibody induced a surface redistribution, it did not, by itself, lead to the release of acrosin, the endpoint of the acrosome reaction. Finally, using the technique of fluorescence recovery after photobleaching, we found that, in the absence of bivalent antibody, mouse sperm surface galactosyltransferase exhibited 40-50% recovery with a high diffusion coefficient on the anterior head (5-8 x 10(-9) cm(2)/s) approximately 2 times greater than on the posterior head (2-4 x 10(-9) cm(2)/s). When galactosyltransferase was cross-linked and redistributed to the anterior head using the bivalent antibody, the mobile fraction decreased to 20-30% with no significant change in the diffusion coefficient. These data demonstrate that mouse sperm surface galactosyltransferase has many important properties requisite for its function as a receptor for the zona pellucida, but bivalent cross-linking of mouse sperm surface galactosyltransferase is not sufficient to induce the final signal transduction events leading acrosomal exocytosis.
机译：已经提出，小鼠的精子表面β-1，4-半乳糖基转移酶在受精过程中用作透明带的受体。在本文中，我们使用了两种对半乳糖基转移酶具有特异性的单价荧光探针：UDP-半乳糖的三硝基苯基化衍生物和Rhodaminatedα-乳清蛋白。我们发现半乳糖基转移酶最初存在于顶体完整精子的后部头部，但在它与抗半乳糖基转移酶多克隆抗体交联后逐渐被定位在覆盖顶体区域的质膜上。用钙离子载体A23187处理的标记的小鼠精子显示，顶体胞吐作用后半乳糖基转移酶保留在后头。但是，如果半乳糖基转移酶先与抗体交联并重新分布，然后顶体与A23187反应，则所有头部荧光都会丢失。另外，尽管抗半乳糖基转移酶抗体诱导了表面再分布，但是它本身并没有导致顶体反应的终点-顶肽释放。最后，使用光漂白后的荧光恢复技术，我们发现，在没有二价抗体的情况下，小鼠精子表面半乳糖基转移酶表现出40-50％的恢复率，并在前额头上具有高扩散系数（5-8 x 10（-9 ）cm（2）/ s）大约比后头（2-4 x 10（-9）cm（2）/ s）大2倍。当使用二价抗体将半乳糖基转移酶交联并重新分配至前脑时，可动部分降低至20-30％，而扩散系数无明显变化。这些数据表明，小鼠精子表面半乳糖基转移酶具有作为透明带的受体所必需的许多重要特性，但是小鼠精子表面半乳糖基转移酶的二价交联不足以诱导导致最终的顶体胞吐作用的最终信号转导事件。

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