Properties of flagellar "rigor waves" formed by abrupt removal of adenosine triphosphate from actively swimming sea urchin sperm.
Properties of flagellar "rigor waves" formed by abrupt removal of adenosine triphosphate from actively swimming sea urchin sperm.
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DOI：
10.1083/jcb.63.3.970
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发表时间：
1974-12
期刊：
影响因子：
--
通讯作者：
Gibbons IR
中科院分区：
文献类型：
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作者：
Gibbons BH;Gibbons IR
将海胆精子脱膜并用含有 0.04% Triton X-100 和 0.03 mM ATP 的溶液重新激活。然后通过将精子悬浮液稀释50倍到不含ATP的再激活溶液中来突然降低ATP浓度。稀释悬浮液中精子的鞭毛不活动，但它们弯曲成各种静止的严格波形，与正常运动期间鞭毛弯曲周期的不同阶段发生的波形非常相似。在二硫苏糖醇和 EDTA 存在下储存几个小时后，这些严酷波的形式没有改变。添加 1 µM ATP 会导致波缓慢松弛，大多数精子在大约 30 分钟的时间内部分变直；浓度稍高一些，放松会更快、更彻底。 ATP 浓度高于 10 µM 会诱导恢复正常跳动运动。添加 ITP、GTP 或 GDP（最多 1 mM）不会导致严酷波松弛。用胰蛋白酶消化至足以破坏径向辐条和连接蛋白连接的程度不会引起严格波形的变化，这表明这些波形可以通过鞭毛轴丝的外双联小管之间的动力蛋白跨桥来维持。粘性剪切对刚性波轴丝影响的研究表明，尽管它们相对容易扭曲，但它们能够抵抗弯曲变形。
Sea urchin sperm were demembranated and reactivated with a solution containing 0.04% Triton X-100 and 0.03 mM ATP. The ATP concentration was then lowered abruptly by diluting the sperm suspension 50-fold into reactivating solution containing no ATP. The flagella of the sperm in the diluted suspension were not motile, but they were bent into a variety of stationary rigor wave forms closely resembling the wave forms occurring at different stages of the flagellar bending cycle during normal movement. The form of these rigor waves was unchanged upon storage for several hours in the presence of dithiothreitol and EDTA. Addition of 1 µM ATP induced slow relaxation of the waves, with most of the sperm becoming partially straightened over a period of about 30 min; somewhat higher concentrations gave a more rapid and complete relaxation. Concentrations of ATP above 10 µM induced resumption of normal beating movements. Addition of ITP, GTP, or GDP (up to 1 mM) produced no relaxation of the rigor waves. Digestion with trypsin to an extent sufficient to disrupt the radial spokes and the nexin links caused no change in the rigor wave forms, suggesting that these wave forms could be maintained by the dynein cross-bridges between the outer doublet tubules of the flagellar axoneme. Study of the effects of viscous shear on the rigor wave axonemes has shown that they are resistant to distortion by bending, although they can be twisted relatively easily.