Interplay between Convective and Viscoelastic Forces Controls the Morphology of In Vitro Paclitaxel-Stabilized Microtubules

Abstract

Microtubules (MTs) are self-assembling, high-aspect-ratio tubular nanostructures formed from the polymerization of tubulin protein. MTs are capable of globally assembling into optically birefringent morphologies, but there is disagreement on the mechanisms driving this behavior. We investigated the temporal evolution of paclitaxel (PTX)-stabilized MT solutions under a range of in vitro conditions. Significant morphological differences were observed in the polymerized PTX-MT solutions as a consequence of varying the orientation of the reaction vessel (vertical vs. horizontal), the type of heating source (hot plate vs. incubator), the incubation time, and the concentration of PTX (high vs. low). The most robust birefringent patterns were found only in vertically oriented cuvettes that were heated asymmetrically on a hot plate, suggesting dependence upon a convective flow, which we confirmed with a combination of optical and thermal imaging. Higher concentrations of PTX led to denser PTX-MT domain formation and brighter birefringence, due to more complete polymerization. Combining our experimental observations, we conclude that birefringent patterns arise principally through a combination of convective and viscoelastic forces, and we identify the sequence of dynamical stages through which they evolve.