Centriole Number and the Accumulation of Microtubules Modulate the Timing of Apical Insertion during Radial Intercalation

Centrioles are microtubule (MT)-based structures that provide important functions during cell migration, cell division, and cell signaling [1]. Modulating centriole number in 3D cell cultures has been shown to influence protrusive behavior [2-5]. Here, we address in vivo the role of centrioles and the accumulation of MTs on the protrusive behavior required during the initiation of radial intercalation. Radial intercalation is an important developmental process whereby cells undergo polarized movements and interdigitate into a more superficial layer [6, 7]. It is commonly employed during metamorphic events, such as the tissue thinning coupled with expansion or during the introduction of different cell types into an epithelium. During radial intercalation, cells emerge from a basal layer by undergoing a process of apical migration, apical insertion, and expansion [8]. In Xenopus skin, multiciliated cells (MCCs), which contain ∼150 centrioles, and ionocytes (ICs), which contain two centrioles, differentiate during the same developmental window, but MCCs complete intercalation prior to ICs. Here, we utilize this difference in timing to create a quantifiable assay for insertion and find that the timing of insertion is modulated by changes in centriole number and the accumulation of acetylated MTs. Additionally, centrioles align between the nucleus and the leading edge creating an axis of migration with apically oriented (+) ends. Using the MT (-) end protein CAMSAP1 fused to the apically positioned Par6 protein, we have artificially reversed the orientation of MTs and find that the accumulation of MTs in either orientation is sufficient to promote apical insertion.