Analysis of PCP Defects in Mammalian Eye Lens

Multicellular tissues and organs often show planar cell polarity (PCP) where the constituent cells align along an axis to form coordinated patterns. Mammalian eye lenses are mainly comprised of epithelial-derived fibre cells, which exhibit highly ordered alignment that is regulated by PCP signaling. Each fibre cell has an apically situated primary cilium and in most cases this is polarized towards the lens anterior pole. Here we describe how to visualize the global cellular alignment of lens fibre cells by examining the suture pattern that is formed by the tips of fibres meeting at the anterior pole. We also describe a method for whole mount preparation, which allows observation of the polarized distribution of primary cilia at the apical surface of lens fibres. Given its relative simplicity, at least in cellular terms, and its requirement for a high degree of precision in cellular alignment and orientation, we predict that the lens will be an excellent model system to help elucidate the role of cilia and PCP components in the development of three-dimensional organization in tissues and organs.