Abstract
To study the properties of tracheal cilia beating under various conditions, we developed a method to monitor the movement of the ciliary tip. One end of a demembranated cilium was immobilized on the glass surface, while the other end was capped with a polystyrene bead and tracked in three dimensions. The cilium, when activated by ATP, stably repeated asymmetric beating as in vivo. The tip of a cilium in effective and recovery strokes moved in discrete trajectories that differed in height. The trajectory remained asymmetric in highly viscous solutions. Model calculation showed that cilia maintained a constant net flux during one beat cycle irrespective of the medium viscosity. When the bead attached to the end was trapped with optical tweezers, it came to display linear oscillation only in the longitudinal direction. Such a beating-mode transition may be an inherent nature of movement-restricted cilia.
Highlights
Motile cilia of eukaryotic cells propagate bending waves and produce water flow over the cell surface
We found that the cilium tip maintained a difference in height between the effective and recovery strokes under various conditions
We developed a method to track the tip of a single beating cilium based on our finding that sulfate-modified fluorescent polystyrene bead spontaneously and predominantly attach to ciliary tips
Summary
Motile cilia of eukaryotic cells propagate bending waves and produce water flow over the cell surface. To further examine the robustness that maintains a constant z-gap, we combined this system with optical trapping of the bead and analyzed the load-dependent behavior of cilia This method enables precise position-control and observation of ciliary tip, as well as application of external force. While periodic oscillation persisted under bead-trapped conditions, the beating trajectory of the ciliary tip changed into a linear one moving only in the z-direction. This oscillation was composed of two different phases, which most likely corresponded to www.nature.com/scientificreports/. 6 μm the effective and recovery strokes in beating cilia, no significant curvature change was observed in the laser-trapped axonemes This behavior indicates a robustness of cilia in beating in distinct effective and recovery phases even under extremely restrictive conditions
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