Alternate patterns of doublet microtubule sliding in ATP-disintegrated macrocilia of the ctenophore Beroë.

Abstract

We have used the unique properties of macrocilia from the lips of the ctenophore Beroë to test whether the ciliary beat cycle is caused by sequential activation of doublet sliding on opposite sides of the axoneme (Satir, P., 1982, Soc. Exp. Biol. Symp., 35: 179-201; Sugino, K., and Y. Naitoh, 1982, Nature (Lond.), 295: 609-611; Wais-Steider, J., and P. Satir, 1979, J. Supramol. Struct., 11:339-347). Macrocilia contain several hundred axonemes linked into rows by lamellae between doublets 3 and 8. These connections provide morphological markers for numbering the doublet microtubules in thin sections. Demembranated, detached macrocilia undergo ATP-induced sliding disintegration by extrusion of thick fragments and finer fibers from the proximal end. Disintegration can easily be followed with low-magnification brightfield or phase-contrast optics. Sliding occurs with or without added elastase, and is reversibly inhibited by vanadate. Thin sections through 16 ATP-disintegrated macrocilia showed two mutually exclusive patterns of doublet extrusion with equal frequency. Doublets 9, 1, and 2 or doublets 5, 6, and 7 were usually extruded, but not both groups. We conclude that both subsets of doublets slide by their own active arms, and that the two extrusion patterns represent alternate activation and inactivation of doublet sliding on opposite halves of the axoneme. These findings provide the first direct experimental support for a switching mechanism regulating microtubule sliding in cilia.