Abstract

Sperm glycocalyx and plasma membrane undergo outstanding modifications during fertilization. However, it is unclear how in vitro capacitation time and acrosome reaction affect the specific location of boar sperm glycoconjugates. This study aimed to identify lectin binding patterns and to describe the sequential changes during different in vitro capacitation times (1 and 4 h) and acrosome reaction in boar spermatozoa. With Aleuria aurantia agglutinin (AAA), most uncapacitated cells were labelled in the postacrosomal region. Nevertheless, after 1 h of in vitro capacitation and the acrosome reaction, most AAA binding sites were in the acrosomal region. With Concanavalin A (ConA), most sperm were labeled in the postacrosomal region before and after capacitation. After the acrosome reaction induction, this pattern changed to a highly stained acrosomal and postacrosomal regions. Peanut agglutinin (PNA) binding sites were in the acrosomal region in uncapacitated and capacitated sperm. In acrosome reacted sperm after 4h capacitation, the most frequent pattern showed remaining positive labeling in the central area of the head. With Pisum sativum agglutinin (PSA), most uncapacitated cells showed a postacrosomal region staining. Nevertheless, faint stained all over the head and highly acrosomal region labelling was observed in the major part of capacitated and acrosome reacted sperm respectively. With Wheat germ agglutinin (WGA), the most representative pattern in uncapacitated, capacitated and acrosome reacted sperm was labelled in the acrosomal region. Regarding capacitation time, the most significant changes in the most representative pattern were observed in acrosome reacted spermatozoa after 4 h of in vitro capacitation.

• Highlights

• Lectins allow detailed descriptions of eight different populations in boar sperm.

• Changes in lectin binding patterns during capacitation are time dependent.

• Lectin binding patterns significantly change after 4 h of in vitro capacitation.

摘要

精子糖萼和质膜在受精过程中会发生明显的改变。但是，尚不清楚体外获能时间和顶体反应如何影响公猪精子糖缀合物的特定位置。这项研究旨在鉴定凝集素的结合模式，并描述在不同的体外获能时间（1和4小时）和公猪精子顶体反应期间的顺序变化。有了Aleuria aurantia凝集素（AAA），大多数未失能的细胞都被标记在了顶体后区域。不过，在体外1小时后获能和顶体反应，大多数AAA结合位点位于顶体区域。使用伴刀豆球蛋白A（ConA），在获能前后，大多数精子都在顶体后区域标记。在顶体反应诱导之后，该模式改变为高度染色的顶体和顶体后区域。花生凝集素（PNA）结合位点位于无能力和有能力的精子的顶体区域。获能4小时后，顶体反应的精子中，最常见的模式显示头部中央区域仍保留阳性标记。与豌豆凝集素（PSA），大多数未失能的细胞显示了顶体后区域的染色。然而，在整个头顶上均出现了微弱的染色，并且在获能的顶体和顶体反应的精子的大部分中都观察到了高度的顶体区域标记。使用小麦胚芽凝集素（WGA），在顶体区域标记了无能力，有能力和顶体反应的精子中最具代表性的模式。关于获能时间，在体外获能4小时后，在顶体反应的精子中观察到最具代表性模式的最显着变化。

• 强调

• 凝集素可以详细描述公猪精子中的八个不同种群。

• 获能期间凝集素结合模式的变化与时间有关。

• 体外获能4小时后，凝集素结合模式发生了显着变化。