The time course of structural changes in vascular smooth muscle cells (SMC) was investigated during the formation of an experimental lesion in response to balloon injury. We compared the filamentous organization, evaluated by quantitative electron microscopy, with the cellular content of two representative cytocontractile proteins (myosin and tropomyosin) as assessed by immunofluorescence. We found that the changes peak between 7 and 14 days after injury and that they are visible both in the neointima and to a lesser extent in the inner media. While virtually all SMC are of a filament-rich phenotype in the undisturbed media, after balloon injury SMC migrated into the intima and about 90% of these latter cells were either of a organelle-rich or an intermediate phenotype, with the remaining 10% being of the filament-rich phenotype. In the inner media about 40% of cells were either of organelle-rich or intermediate phenotype. In contrast to these profound organizational changes of responding SMC, histochemistry revealed only a slight and probably transient decrease of the cellular content of myosin and tropomyosin at that time point. Twenty-eight days after injury the discrepancies between the content and the organization of cytocontractile proteins became more apparent. While virtually all SMC showed a homogeneous intensive staining with both antibodies, indistinguishable from the media SMC, the organization of cytoplasmic filaments had not totally recovered. Even though this morphological study does not permit conclusions to be drawn on the contractile function of the cells, it shows that both the organization and the content of cytocontractile protein have to be analyzed and compared for SMC changes to be evaluated during the formation of an experimental lesion.