Flagellar arrest behavior predicted by the geometric clutch model is confirmed experimentally by micromanipulation experiments on reactivated bull sperm

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Abstract
The central tenet of the Geometric Clutch hypothesis of flagellar beating is that the internal force transverse to the outer doublets (t-force) mediates the initiation and termination of episodes of dynein engagement. Therefore, if the development of an adequate t-force is prevented, then the dynein-switching necessary to complete a cycle of beating should fail. The dominant component of the t-force is the product of the longitudinal force on each outer doublet multiplied by the local curvature of the flagellum. In the present study, two separate strategies, blocking and clipping, were employed to limit the development of the t-force in Triton X-100 extracted bull sperm models. The blocking strategy used a bent glass microprobe to restrict the flagellum during a beat, preventing the development of curvature in the basal portion of the flagellum. The clipping strategy was designed to shorten the flagellum by clipping off distal segments of the flagellum with a glass microprobe. This limits the number of dyneins that can contribute to bending and consequently reduces the longitudinal force on the doublets. The blocking and clipping strategies both produced an arrest of the beat cycle consistent with predictions based on the Geometric Clutch hypothesis. Direct comparison of experimentally produced arrest behavior to the behavior of the Geometric Clutch computer model of a bull sperm yielded similar arrest patterns. The computer model duplicated the observed behavior using reasonable values for dynein force and flagellar stiffness. The experimental data derived from both blocking and clipping experiments are fully compatible with the Geometric Clutch hypothesis.Cell Motil. Cytoskeleton 44:177–189, 1999.