Italiano J E, Stewart M, Roberts T M
Department of Biological Science, Florida State University, Tallahassee, Florida 32306, USA.
J Cell Biol. 1999 Sep 6;146(5):1087-96. doi: 10.1083/jcb.146.5.1087.
The major sperm protein (MSP)-based amoeboid motility of Ascaris suum sperm requires coordinated lamellipodial protrusion and cell body retraction. In these cells, protrusion and retraction are tightly coupled to the assembly and disassembly of the cytoskeleton at opposite ends of the lamellipodium. Although polymerization along the leading edge appears to drive protrusion, the behavior of sperm tethered to the substrate showed that an additional force is required to pull the cell body forward. To examine the mechanism of cell body movement, we used pH to uncouple cytoskeletal polymerization and depolymerization. In sperm treated with pH 6.75 buffer, protrusion of the leading edge slowed dramatically while both cytoskeletal disassembly at the base of the lamellipodium and cell body retraction continued. At pH 6.35, the cytoskeleton pulled away from the leading edge and receded through the lamellipodium as its disassembly at the cell body continued. The cytoskeleton disassembled rapidly and completely in cells treated at pH 5.5, but reformed when the cells were washed with physiological buffer. Cytoskeletal reassembly occurred at the lamellipodial margin and caused membrane protrusion, but the cell body did not move until the cytoskeleton was rebuilt and depolymerization resumed. These results indicate that cell body retraction is mediated by tension in the cytoskeleton, correlated with MSP depolymerization at the base of the lamellipodium.
猪蛔虫精子基于主要精子蛋白(MSP)的阿米巴样运动需要片状伪足的突出和细胞体的收缩协调进行。在这些细胞中,突出和收缩与片状伪足两端细胞骨架的组装和拆卸紧密相关。虽然沿着前缘的聚合作用似乎驱动了突出,但附着在底物上的精子的行为表明,需要额外的力来向前拉动细胞体。为了研究细胞体运动的机制,我们利用pH值来解偶联细胞骨架的聚合和解聚。在用pH 6.75缓冲液处理的精子中,前缘的突出显著减慢,而片状伪足基部的细胞骨架拆卸和细胞体收缩仍在继续。在pH 6.35时,细胞骨架从前缘拉开,并随着其在细胞体处的拆卸继续通过片状伪足后退。在pH 5.5处理的细胞中,细胞骨架迅速且完全地拆卸,但当用生理缓冲液洗涤细胞时又重新形成。细胞骨架在片状伪足边缘重新组装并导致膜突出,但直到细胞骨架重建且解聚恢复时细胞体才移动。这些结果表明,细胞体收缩是由细胞骨架中的张力介导的,这与片状伪足基部的MSP解聚相关。