Effects of osmotic force and torque on microtubule bundling and pattern formation

We report effects of polyethylene glycol (PEG, molecular weight of 35 kDa ) on microtubule (MT) bundling and pattern formation. Without PEG, polymerizing tubulin solutions of a few mg/ml that are initially subjected to a field that aligns MTs can spontaneously form striated birefringence patterns. These patterns form through MT alignment, bundling, and coordinated bundle buckling. With increasing PEG concentrations, solutions form progressively weaker patterns. At a sufficiently high PEG concentration ( approximately 0.5% by weight), the samples maintain a nearly uniform birefringence (i.e., no pattern) and laterally contract at a later stage. Concomitantly, on a microscopic level, the network of dispersed MTs that accompany the bundles in pure solutions disappear and the bundles become more distinct. We attribute the weakening of the pattern to the loss of the dispersed MT network, which is required to mediate the coordination of bundle buckling. We propose that the loss of the dispersed network and the enhanced bundling result from PEG associated osmotic forces that drive MTs together and osmotic torques that facilitate their bundling. Similarly, we attribute the lateral contraction of the samples to osmotic torques that tend to align crossing bundles in the network.