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利用场发射扫描电子显微镜(FE-SEM)对人类精子中末端和连接型岩藻糖残基进行定量及拓扑分布研究

Quantification and Topographical Distribution of Terminal and Linked Fucose Residues in Human Spermatozoa by Using Field Emission Scanning Electron Microscopy (FE-SEM).

作者信息

Robles-Gómez Laura, Sáez-Espinosa Paula, López-Viloria Eliana Marina, López-Botella Andrea, Aizpurua Jon, Gómez-Torres María José

机构信息

Departamento de Biotecnología, Universidad de Alicante, 03080 Alicante, Spain.

IVF Spain, Reproductive Medicine, 03540 Alicante, Spain.

出版信息

Int J Mol Sci. 2021 Nov 4;22(21):11947. doi: 10.3390/ijms222111947.

Abstract

The modification of sperm glycocalyx is an essential process during sperm capacitation. The presence and redistribution of terminal and linked fucose have been described during in vitro capacitation in humans. However, the influence of the capacitation time on the quantification and localization of terminal and linked fucose is still unknown. In this study, the quantitative and qualitative changes in fucosyl residues during different in vitro capacitation times (1 and 4 h), are simultaneously characterized by using (AAA) lectin-gold labelling and high-resolution field emission scanning electron microscopy (FE-SEM) in human sperm. A significant decrease was found in the number of terminal fucose registered in the whole sperm head during the in vitro capacitation. Nevertheless, the quantification of fucose residues after 1 h of in vitro capacitation was very similar to those found after 4 h. Therefore, the changes observed in terminal and linked fucose during capacitation were not time-dependent. Furthermore, the comprehensive analysis of the topographic distribution showed the preferential fucosyl location in the acrosomal region and the presence of distinct clusters distributed over the head in all the studied conditions. Overall, these findings corroborate the validity of FE-SEM combined with gold labelling to register changes in surface molecules during in vitro sperm capacitation.

摘要

精子糖萼的修饰是精子获能过程中的一个重要环节。在人类体外获能过程中,已对末端和连接型岩藻糖的存在及重新分布进行了描述。然而,获能时间对末端和连接型岩藻糖的定量及定位的影响仍不清楚。在本研究中,通过使用(AAA)凝集素-金标记和高分辨率场发射扫描电子显微镜(FE-SEM),同时对人类精子在不同体外获能时间(1小时和4小时)内岩藻糖基残基的定量和定性变化进行了表征。在体外获能过程中,整个精子头部记录的末端岩藻糖数量显著减少。然而,体外获能1小时后岩藻糖残基的定量与4小时后发现的情况非常相似。因此,获能过程中末端和连接型岩藻糖的变化不是时间依赖性的。此外,对地形分布的综合分析表明,在所有研究条件下,岩藻糖基优先位于顶体区域,并且在头部存在不同的簇。总体而言,这些发现证实了FE-SEM结合金标记在记录体外精子获能过程中表面分子变化方面的有效性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae00/8584901/eb3dc1c7fe1b/ijms-22-11947-g001.jpg

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