Determinants of cytoplasmic microtubule depolymerization during ciliogenesis in Chlamydomonas

Abstract

At the core of cilia are microtubules which establish length and assist ciliary assembly and disassembly; however, microtubules outside of the cilium can regulate ciliogenesis. The microtubule cytoskeleton polymerizes and depolymerizes rapidly. These processes have been studied across various organisms with chemical and genetic perturbations. However, these have generated con-flicting data in terms of the role of cytoplasmic microtubules (CytoMTs) and free tubulin dynamics during ciliogenesis. Here, we look at the relationship between ciliogenesis and CytoMT dynamics in Chlamydomonas reinhardtii using chemical and mechanical perturbations. We find that not only can stabilized CytoMTs allow for normal ciliary assembly, but high calcium concentrations and low pH-induced deciliation cause CytoMTs to depolymerize separately from ciliary shedding. In addition, ciliary shedding through mechanical shearing allows cilia to regenerate earlier despite intact CytoMTs. Our data suggest that CytoMTs are not a sink for a limiting pool of cytoplasmic tubulin in Chlamydomonas, depolymerization after deciliation is a consequence rather than a requirement for ciliogenesis, and intact tubulin in the cytoplasm and proximal cilium support more efficient ciliary assembly.