Lectin staining and flow cytometry reveals female-induced sperm acrosome reaction and surface carbohydrate reorganization.
Lectin staining and flow cytometry reveals female-induced sperm acrosome reaction and surface carbohydrate reorganization.
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DOI：
10.1038/srep15321
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发表时间：
2015-10-16
影响因子：
4.6
通讯作者：
Evans JP
中科院分区：
文献类型：
--
作者：
Kekäläinen J;Larma I;Linden M;Evans JP
所有细胞都被聚糖覆盖，这是一种独特的寡糖和多糖层，是自我识别和其他细胞水平相互作用（例如受精）的关键调节剂。这些过程之间的功能相似性表明，配子表面聚糖也可能有一个重要的，但目前被忽视，在性选择的作用。在这里，我们开发了一个用户友好的方法，旨在促进未来的测试这种可能性。我们提出的方法是基于流式细胞术定量的女性诱导的精子顶体反应和精子表面聚糖修饰的地中海贻贝。在这个物种中，与许多其他类群一样，卵子释放水溶性因子，吸引同种精子（化学吸引力），并促进精子行为和生理学的潜在可测量变化。我们证明，流式细胞术能够识别精子从其他海水颗粒，以及准确地测量顶体反应和精子聚糖的结构修饰。这种方法可以增加我们的理解化学调节配子水平的相互作用和个体特异性配子识别贻贝和其他类群具有相似的，易于识别的顶体结构。我们的方法也可能适用于其他几个物种，因为碳水化合物介导的细胞水平的配子之间的相互作用是普遍的外部和内部的物种。
All cells are covered by glycans, an individually unique layer of oligo- and polysaccharides that are critical moderators of self-recognition and other cellular-level interactions (e.g. fertilization). The functional similarity between these processes suggests that gamete surface glycans may also have an important, but currently overlooked, role in sexual selection. Here we develop a user-friendly methodological approach designed to facilitate future tests of this possibility. Our proposed method is based on flow cytometric quantification of female-induced sperm acrosome reaction and sperm surface glycan modifications in the Mediterranean mussel Mytilus galloprovincialis. In this species, as with many other taxa, eggs release water-soluble factors that attract conspecific sperm (chemoattraction) and promote potentially measurable changes in sperm behavior and physiology. We demonstrate that flow cytometry is able to identify sperm from other seawater particles as well as accurately measure both acrosome reaction and structural modifications in sperm glycans. This methodological approach can increase our understanding of chemically-moderated gamete-level interactions and individual-specific gamete recognition in Mytilus sp. and other taxa with similar, easily identifiable acrosome structure. Our approach is also likely to be applicable to several other species, since carbohydrate-mediated cellular-level interactions between gametes are universal among externally and internally fertilizing species.