Investigating Localization of Prickle Homologs in the Mammalian Epithelial Cells

Abstract

Planar cell polarity (PCP), or the property that accounts for the orientation andalignment of cells within the plane of an epithelial sheet, is dependent on a network ofPCP proteins that transmit the signals needed to coordinate the local and globalpolarization of cells. PCP has been predominantly studied in the Drosophila wing, and inthe mammalian hair follicles. Prickle is one core PCP protein that has been associatedwith the transmembrane PCP protein known as Vangl2, on the proximal/anterior side ofcells. Three Prickle homologs have been identified in mammals; however, theirlocalization in the basal layer has not yet been assayed. We have generated GFP reporterconstructs for Prickle 1, Prickle 2, and Prickle 3 under the CAG promoter to driveexpression of each Prickle homolog. We show here that there are significant differencesin localization for each GFP-tagged Prickle protein, suggesting possible unique functionsthat have not yet been identified for the three Prickle homologs. We also show thatPrickle2-GFP localizes to cell junctions when Celsr1 is exogenously introduced incultured skin cells. Finally, we propose utilizing the Prickle-GFP constructs forgenerating transgenic embryos via electroporation. This method would allow for earlierscreening and higher throughput of gene transfer than the commonly used methods fortransgenesis. This study focused on investigating the localization of three understudied,but essential, PCP components in order to gain further understanding of PCP proteininteractions.