Analysis of the effect of various PKA and ciliary modulators on the migration of RPE cells

Abstract

Cellular migration is a common activity for many different types of cells. If cellular migration is disrupted, cells may be unable to repair damaged tissue (wound repair), migrate appropriately, or correctly signal their cellular state. In several situations, primary cilia have been implicated in helping to determine the direction of cellular migration. cAMP-dependent Protein Kinase A (PKA) regulates many of the proteins involved in the migration process, but its effects vary with cell types. PKA is also known to localize in primary cilia. Using Matrigel invasion chambers as a 3D model of cellular migration, various PKA and ciliary reagents were tested for their ability to alter retinal pigment epithelial (RPE) cell motility. We used ARPE.19 cells, a human RPE cell line capable of forming cilia in culture, as the cell model. Our results showed that 8-Br-cAMP, LiCl and H89 significantly inhibited migration, while bFGF and NaCl promoted migration in ARPE.19 cells. Forskolin was significant as well in reducing migration. While KT5720 reduced migration, it was not significant, although it appears to be trending that way. These results suggest that PKA may be a key link between ciliary signaling and the direction of cellular migration.