Polarized microtubule gliding and particle saltations produced by soluble factors from sea urchin eggs and embryos.
In this report, we describe an in vitro system for analyzing microtubule-based movements in supernatants of sea urchin egg and embryo homogenates. Using video enhanced DIC microscopy, we have observed bidirectional saltatory particle movements on native taxol-stabilized microtubules assembled in low speed supernatants of Lytechinus egg homogenates, and gliding of these microtubules across a glass surface. A high speed supernatant of soluble proteins, depleted of organelles, microtubules, and their associated proteins supports the gliding of exogenous microtubules and translocation of polystyrene beads along these microtubules. The direction of microtubule gliding has been determined directly by observation of the gliding of flagellar axonemes in which the (+) and (-) ends could be distinguished by biased polar growth of microtubules off the ends. Microtubule gliding is toward the (-) end of the microtubule, is ATP sensitive, and inhibited only by high concentrations of vanadate. These characteristics suggest that the transport complex responsible for microtubule gliding in S2 is kinesin-like. The implications of these molecular interactions for mitosis and other motile events are discussed.
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000502:physiology*; D005260:Female; D008857:Microscopy, Interference / Q000379:methods; D008870:Microtubules / Q000187:drug effects / Q000648:ultrastructure*; D010063:Ovum / Q000166:cytology / Q000502:physiology*; D017239:Paclitaxel; D012617:Sea Urchins / Q000196:embryology*
D000470:Alkaloids; D017239:Paclitaxel