Related Experiment Video
Updated: Aug 24, 2026

Methods to Study Mrp4-containing Macromolecular Complexes in the Regulation of Fibroblast Migration
Published on: May 19, 2016
PDE4-regulated cAMP degradation controls the assembly of integrin-dependent actin adhesion structures and REF52 cell
Yvonne M Fleming1, Margaret C Frame, Miles D Houslay
1Institute of Biological and Life Sciences, Davidson and Wolfson Buildings, University of Glasgow, Glasgow G12 8QQ, UK. y.fleming@beatson.gla.ac.uk
Abstract:
Plating of REF52 cells onto extracellular matrix components leads to the formation of integrin-dependent actin adhesion microspikes. We show that the formation of these structures is sensitive to chemical agents that regulate cAMP levels, such as forskolin and IBMX. In particular, by using the specific inhibitor rolipram, we identify the PDE4 family of cAMP-specific phosphodiesterases as critical regulators of this process. The effect of PDE4 on microspike formation is mediated by actions exerted through the activation of PKA - rather than through the alternative cAMP effector, Epac. We provide evidence that peripheral microspikes are RhoA-, ROCK- and myosin-dependent, and that this pathway is suppressed by PDE4 inhibition. In addition, PDE4 inhibition impairs cell locomotion that requires dynamic protrusion and retraction of peripheral spike structures. Our data demonstrate that PDE4 activity is a key modulator of integrin-induced actin assembly at the cell periphery which, in turn, controls cell migration.
Related Concept Videos
Intracellular Signaling Affects Focal Adhesions
Some...
Mechanism of Lamellipodia Formation
Mechanism of Filopodia Formation
Their main function is to guide migrating cells during normal tissue morphogenesis or cancer metastasis by recognizing and making initial contacts with the extracellular matrix. However, they can also act as stationary cell anchors or help to establish communication...
Activation of Integrins
In "outside-in signaling," external factors in the extracellular space bind to exposed ligand binding sites on integrins. This causes the inactive protein to undergo a conformational change to become active. Integrins are often clustered on the cell membrane. Repetitive and regularly spaced ligand binding events provide an effective stimulus.
Cytoskeletal Coordination in Cell Migration
Actin Polymerization and Cell Motility
Actin cytoskeleton dynamics can produce pushing, pulling, and resistance forces that help the cell to migrate.

