Roberts T M, Ward S
J Cell Biol. 1982 Jan;92(1):132-8. doi: 10.1083/jcb.92.1.132.
Latex beads and wheat germ agglutinin (WGA) were used to examine the movement of membrane components on amoeboid spermatozoa of Caenorhabditis elegans. The behavior of beads attached to the cell revealed continuous, directed movement from the tip of the pseudopod to its base, but no movement on the cell body. Lectin receptors are also cleared from the pseudopod (4). Blocking preexisting lectin receptors with unlabeled WGA followed by pulse-labeling wih fluorescent WGA showed that new lectin receptors are continuously inserted at the tip of the pseudopod. Like latex beads, these new lectin receptors move continuously over the pseudopod surface to the cell body-pseudopod junction where they are probably internalized. Mutants altering the rate of membrane flow, and eliminating its topographical asymmetry, have been identified. Together with the observation that fluorescent phospholipids are cleared from the pseudopod of developing spermatozoa at the same rate as lectin receptors (25), these results show that there is bulk membrane flow over the pseudopod with assembly at the tip and apparent disassembly at the base. There are no vesicles visible at either the pseudopodial tip or base, so these spermatozoa must have a novel mechanism for insertion and uptake of membrane components. This membrane flow could provide the forward propulsion of spermatozoa attached to a substrate by their pseudopods.
利用乳胶珠和小麦胚凝集素(WGA)来检测秀丽隐杆线虫变形虫状精子上膜成分的移动。附着在细胞上的珠子的行为显示,其从伪足尖端向基部持续、定向移动,但在细胞体上没有移动。凝集素受体也从伪足上清除(4)。用未标记的WGA阻断预先存在的凝集素受体,然后用荧光WGA进行脉冲标记,结果表明新的凝集素受体在伪足尖端持续插入。与乳胶珠一样,这些新的凝集素受体在伪足表面持续移动到细胞体 - 伪足连接处,在那里它们可能被内化。已经鉴定出改变膜流动速率并消除其拓扑不对称性的突变体。再加上观察到发育中的精子伪足中的荧光磷脂与凝集素受体以相同速率清除(25),这些结果表明在伪足上存在大量膜流动,在尖端组装,在基部明显拆卸。在伪足尖端或基部均未观察到囊泡,因此这些精子必定具有一种用于膜成分插入和摄取的新机制。这种膜流动可以为由伪足附着在底物上的精子提供向前推进力。