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

通过测量外双微管之间的滑动来分析金仓鼠精子的鞭毛运动

基本信息

批准号：
02640551
负责人：
ISHIJIMA Sumio
金额：
$ 0.96万
依托单位：
Tokyo Institute of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02640551/
关键词：
Flagellar movement Hamster Microtubule Sliding Sperm

项目摘要

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.

1.在金黄地鼠精子中建立了“鞭毛模型”的方法，即脱膜精子在MgATP_（12）激活时显示出与完整精子相似的鞭毛运动<2->。精子绕其长轴的滚动运动和精子在盖玻片下的偏转运动的方向以及鞭毛波的振幅和不对称性都受再激活液中Ca^2+浓度的影响。与海胆精子相反，大多数完整的金黄地鼠精子从头部观察时呈逆时针方向滚动。因此，金黄地鼠精子细胞内Ca^<2+>浓度可能与海胆精子细胞内Ca^<2+>浓度不同.用DTT和胰蛋白酶处理脱膜精子，然后加入MgATP-2<2->，在暗视野显微镜下观察到外双微管从轴丝上滑动。通过测量微管的长度和时间确定的这种滑动的速度约为8 μ m/s。介质中Ca^2+浓度对滑动速度的影响不大.睾丸精子与附睾头、尾精子的滑动速度无显著性差异。这些结果证实，这些精子中的运动装置已经在睾丸中功能性组装，尽管这些精子在台氏液中是不动的。