In Situ Hybridization Analysis of Vascular Endothelium

Abstract

In situ hybridization with RNA probes is a valuable technique to determine the expression pattern of endothelial genes in tissue and allows the spatial localization of the corresponding mRNA at the cellular level. Labeled single-stranded complementary RNA probes (cRNA) are used to detect the mRNA of interest on histological sections, i.e., in situ. In comparison to the immunological detection of gene products, in situ hybridization is more time-consuming and has a lower resolution at the cellular level. However, it is the method of choice if suitable antibodies to detect the protein itself are not available. Moreover, the detection of mRNA allows to unambigously determine the site of synthesis of endothelial receptors, of secreted molecules such as angiogenic factors, or of molecules whose extracellular domains can be proteolytically processed, and which, upon release from their producer cells, may travel to distant locations in the tissue (e.g., Breier et al. 1995, Breier et al. 1996, Steffen et al. 1996, Vajkoczy et al. 2002).