Bawa S R, Pabst M A, Werner G, Bains H K
Department of Biophysics, Panjab University, Chandigarh, India.
Andrologia. 1993 May-Jun;25(3):123-35. doi: 10.1111/j.1439-0272.1993.tb02694.x.
Plasma membrane alterations accompanying in vitro capacitation and acrosome reaction of goat spermatozoa were studied using lectin labelling, scanning electron microscopy, and freeze-fracture methods. Fluorescein isothiocyanate linked lectins namely; Canavalia ensiformis (ConA), Maclura pomifera (MPA), Arachis hypogaea (PNA), Glycine max (SBA) and Triticum vulgaris (WGA) agglutinin were used to examine the distribution of surface carbohydrates during these two events. The head and the sperm tail reveal altered lectin labelling features after capacitation and acrosome reaction. After capacitation the surface coat components for MPA, SBA, and WGA are shed from the spermatozoon head. ConA receptors on the head are retained after capacitation but are partially shed in the acrosome reacted spermatozoa. SBA receptor sites appear on the sperm tail of the capacitated spermatozoa. Unusual morphological changes attending capacitation involve the sperm tail-end which develops a novel entity, which we have termed 'spatula'. The 'spatula' shows strong binding with ConA and WGA only. In the acrosome reacted spermatozoa the spatulated tail-end unwinds with a concomitant loss of lectin labelling. Highly ordered membrane particles, 'ladders' of the middle piece of the epididymal sperm tail, disappear and IMP clearings appear on the middle piece and in the spatulated ends of the capacitated spermatozoa. But in the acrosome reacted sperm IMPs reappear and are randomly disposed on the middle-piece and are clustered in small patches on the principal-piece. IMP free areas appear on the plasma membrane covering the acrosome and the outer acrosomal membrane (OAM) of the capacitated spermatozoa. The plasma membrane and OAM fuse at multiple foci and appear as acrosomal 'ghosts' which remain associated with the sperm head even after acrosome reaction.
采用凝集素标记、扫描电子显微镜和冷冻断裂方法,研究了山羊精子体外获能和顶体反应过程中质膜的变化。使用异硫氰酸荧光素标记的凝集素,即刀豆球蛋白A(ConA)、桑科榕属植物(MPA)、花生凝集素(PNA)、大豆凝集素(SBA)和小麦胚凝集素(WGA),来检测这两个过程中表面碳水化合物的分布。精子头部和尾部在获能和顶体反应后显示出凝集素标记特征的改变。获能后,MPA、SBA和WGA的表面 coat 成分从精子头部脱落。头部的ConA受体在获能后保留,但在顶体反应的精子中部分脱落。SBA受体位点出现在获能精子的尾部。获能伴随的异常形态变化涉及精子尾端,尾端形成一个新的结构,我们称之为“刮刀”。“刮刀”仅与ConA和WGA有强结合。在顶体反应的精子中,带刮刀的尾端展开,同时凝集素标记消失。附睾精子尾部中段高度有序的膜颗粒“梯子”消失,在获能精子的中段和带刮刀的尾端出现IMP清除区。但在顶体反应的精子中,IMPs重新出现,并随机分布在中段,在主段聚集成小斑块。在覆盖获能精子顶体和顶体外膜(OAM)的质膜上出现IMP-free区。质膜和OAM在多个位点融合,形成顶体“幽灵”,即使在顶体反应后仍与精子头部相连。