Analysis of flagellar movement of golden hamster spermatozoa by measuring sliding between outer doublet microtubules

1.在金黄地鼠精子中发展了一种“鞭毛模型”的方法，即去膜精子在用镁激活时表现出与完整精子相似的鞭毛运动。复活液中的Ca~(2+)浓度改变了精子绕其长轴的滚动运动方向和精子在盖板下的偏航运动方向以及鞭毛波的幅度和不对称性。与海胆精子相反，从头部看，大多数完整的金黄地鼠精子是逆时针滚动的。因此，金黄地鼠精子胞内Ca~(2+)浓度可能与海胆精子不同。去膜精子用DTT和胰酶处理后，再加入镁，在暗场显微镜下观察到外周微管从轴丝上滑脱。通过测量微管长度和时间确定的滑动速度约为8um/S，滑动速度不受介质中Ca~(2+)浓度的影响。睾丸精子与附睾头、尾部精子的滑动速度无显著差异。这些结果证实，这些精子中的运动器已经在睾丸中进行了功能性组装，尽管这些精子在泰罗氏溶液中是静止的。

1. The method of "flagellar model", which is demembranated spermatozoa showing the flagellar movement similar to that of intact spermatozoa when activated with MgATP^<2->, has been developed in golden hamster spermatozoa. The direction of rolling motion of a spermatozoon about its long axis and of yawing motion of spermatozoa under the coverslip as well as the amplitude and asymmetry of flagellar wave was changed by the Ca^<2+> concentration in reactivation solution. In contrast to the sea urchin spermatozoa, the majority of intact golden hamster spermatozoa rolled counterclockwise when viewed from the head. Thus, intracellular concentration of Ca^<2+> of golden hamster spermatozoa is perhaps different from that of sea urchin spermatozoa.2. When the demembranated spermatozoa were treated with DTT and trypsin, and then MgATP^<2-> was added to them, sliding of the outer doublet microtubules from the axoneme was observed under a darkfield microscope. The velocity of this sliding, which was determined by measuring the length of microtubules and time, was approximately 8 um/s. The sliding velocity did not change by the Ca^<2+> concentration in the medium so much.3. There was no significant difference in the sliding velocity among testicular and caput and cauda epididymal spermatozoa. These results confirmed that the motor apparatus in these spermatozoa is already functionally assembled in the testis, although these spermatozoa are immotile in the Tyrode's solution.