Dedifferentiation of smooth muscle cells (SMC) from the contractile to the synthetic phenotype is a key event in atherosclerosis. A comparable phenotypic change from the contractile to the synthetic state is rapidly incurred when SMC are grown in culture. To identify genes that characterize the contractile and synthetic phenotypes, we performed differential display reverse transcription polymerase chain reactions on RNA from porcine arterial contractile SMC obtained directly from medial tissues and from SMC made synthetic by cell culturing. One of the differentially expressed cDNAs we identified encoded tropomyosin 4 (TM4). Whereas basal levels of TM4 existed in contractile SMC, the amount of TM4 transcripts strongly increased in synthetic SMC (33% vs. 86 – 106% p<0.005). Induction of foam cell formation had no additional enhancing effect on the expression of TM4 in cultivated SMC. We also tested whether TM4 expression was correspondingly enhanced during atherogenesis. The number of TM4-expressing SMC increased with plaque development as demonstrated by simultaneous in situ hybridization and immunohistochemistry. We compared the localization patterns of myosin heavy chain isoforms in normal arteries and lesions of increasing severity and determined that TM4 expression was relegated mainly to SMC of the synthetic phenotype in the media and intima during atherogenesis. The present study demonstrates that upregulation of TM4 mRNA is a relevant marker of dedifferentiation in vascular SMC.