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精子黏附素AQN1是一种参与猪输卵管精子库形成的候选受体分子。

Spermadhesin AQN1 is a candidate receptor molecule involved in the formation of the oviductal sperm reservoir in the pig.

作者信息

Ekhlasi-Hundrieser Mahnaz, Gohr Katrin, Wagner Andrea, Tsolova Miroslava, Petrunkina Anna, Töpfer-Petersen Edda

机构信息

Institute of Reproductive Medicine, School of Veterinary Medicine Hannover, Foundation, 30559 Hannover, Germany.

出版信息

Biol Reprod. 2005 Sep;73(3):536-45. doi: 10.1095/biolreprod.105.040824. Epub 2005 May 11.

Abstract

Sperm are stored in the isthmic region of the oviduct under conditions that maintain viability and suppress early capacitation steps until ovulation occurs. The initial contact between sperm and oviductal epithelium is mediated by carbohydrate-protein interactions. In the pig, the carbohydrate recognition system has been shown to involve oligomannosyl structures. The spermadhesins AWN and AQN1 are the dominant porcine carbohydrate-binding sperm proteins. The objective of this study was to demonstrate that AQN1 contributes to sperm binding to the oviductal epithelium. AQN1 showed a broad carbohydrate-binding pattern as it recognizes both alpha- and beta-linked galactose as well as Manalpha1-3(Manalpha1-6)Man structures, whereas AWN bound only the galactose species. Binding of ejaculated sperm to oviductal epithelium was inhibited by addition of AQN1 but not by AWN. Mannose-binding sites were localized over the rostral region of the sperm head. Flow cytometry showed that, under capacitating conditions, the population of live sperm was shifted within 30 min toward an increase in the proportion of cells with low mannose- and high galactose-binding. The loss of mannose-binding sites was accompanied by the loss of AQN1 in sperm extracts and the significant reduction in the sperm-oviduct binding. The oviductal epithelium was shown by GNA-lectin histochemistry and by SDS-PAGE and lectin blotting of the apical membrane fraction to express mannose components that could be recognized by AQN1. These results demonstrate that the sperm lectin AQN1 fulfils the criteria for an oviduct receptor in the pig and may play a role in the formation of the oviductal sperm reservoir.

摘要

精子储存在输卵管峡部,在此条件下可维持其活力并抑制早期获能步骤,直至排卵发生。精子与输卵管上皮的初始接触是由碳水化合物 - 蛋白质相互作用介导的。在猪中,碳水化合物识别系统已被证明涉及寡甘露糖结构。精子黏附素AWN和AQN1是猪中主要的碳水化合物结合精子蛋白。本研究的目的是证明AQN1有助于精子与输卵管上皮的结合。AQN1表现出广泛的碳水化合物结合模式,因为它既能识别α - 连接和β - 连接的半乳糖,也能识别Manα1 - 3(Manα1 - 6)Man结构,而AWN仅结合半乳糖种类。添加AQN1可抑制射出精子与输卵管上皮的结合,而添加AWN则无此作用。甘露糖结合位点定位于精子头部的吻侧区域。流式细胞术显示,在获能条件下,活精子群体在30分钟内发生变化,低甘露糖结合和高半乳糖结合细胞的比例增加。甘露糖结合位点的丧失伴随着精子提取物中AQN1的丧失以及精子与输卵管结合的显著减少。通过GNA - 凝集素组织化学以及顶端膜部分的SDS - PAGE和凝集素印迹法表明,输卵管上皮表达可被AQN1识别的甘露糖成分。这些结果表明,精子凝集素AQN1符合猪输卵管受体的标准,可能在输卵管精子储存库的形成中发挥作用。

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