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

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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.