Role of myosin light-chain phosphorylation and microtubules in stress fiber morphology in cultured mesangial cells

Abstract

Induced elevations in intracellular cAMP caused remarkable shape changes in cultured mesangial cells that were prevented by prostaglandin E2 (PGE2). The purpose of this study was to determine the morphological and physiological basis for these alterations. Coincident with elevated levels of cAMP and shape change there was rapid dissolution of stress fibers. Cytochalasin B treatment also caused shape change and stress fiber dissolution. Stress fibers were visualized by fluorescence microscopy after appropriate staining. In such cells, microtubules were intact. PGE2 prevented stress fiber loss as well as shape change if it was included in the medium from the outset, or restored stress fibers and normal morphology within 30 min if it was added following shape change. Agents that depolymerize microtubules also prevented both stress fiber loss and shape change. Coincident with stress fiber loss and shape change induced by elevations of cAMP, there was a decrease in the rate of [32P]-orthophosphate incorporation into myosin light chain; this was also prevented or reversed by PGE2. Remarkably, PGE2 alone caused an increase in the rate of [32P]orthophosphate incorporation into myosin light chain in cells that were otherwise untreated. Phosphorylation of the light chain of myosin may be an important factor in the maintenance of stress fiber morphology. The results of this study point out complex interrelationships between microtubules, and stress fibers in the maintenance of cell shape needing further study. The model systems that are presented seem suited for such investigation.