Differential versican isoforms and aggrecan expression in the chicken embryo aorta

Members of the family of large chondroitin sulfate proteoglycans (CSPGs), such as versican and aggrecan, are involved in early heart development, and in the development and progression of atherosclerosis and restenosis. Given the important roles played by versican and aggrecan in such processes, we sought to determine whether these molecules are present in the aortic wall during the advanced stages of chicken embryo development and the endothelial-mesenchymal transformation (EMT). Immunolabeling of serial cryo-sections revealed versican immunoreactivity around the cells within the intimal thickening, and the cells organized in lamellar and interlamellar cell layers. In contrast, a weak aggrecan immunoreactivity was limited to the cells arranged into lamellar and interlamellar cell layers. Immunolabeling also demonstrated that V2 is the main versican isoform present at the intimal thickening. According to immunoblotting analysis, the aggrecan content was very low in all stages examined, and two versican isoforms (V0 and V2) were present at day 14 of development. We also investigated whether versican isoforms were present during EMT in vitro. Versican immunoreactivity was detected in patches of endothelial cells; in the detaching and migrating cells, and the extracellular matrix (ECM) deposited by them; and in cells that had acquired mesenchymal characteristics. These data indicate that versican and aggrecan have different spatial and temporal patterns of expression, and they have different functions during remodeling of the aortic wall. Also, the different immunoreactivity and immunolocalization patterns observed for versican both in vivo and in vitro, in addition to being associated with the presence of different versican isoforms, may be related to the predominance of the V2 isoform during intimal thickening formation and EMT. (C) 2004 Wiley-Liss, Inc.