mRNA Detection in Cerebral Vessels by Nonradioactive In Situ Hybridization

In situ hybridization (ISH) has become a critical tool for studying gene expression in the central nervous system (CNS) (1 –6 ). ISH has advantages over immunohisto-chemistry, because ISH identifies the cells that make the antigen of interest, rather than just contain it. ISH has been used to assess the cellular distribution of messenger ribonucleic acids (mRNAs) encoding many important proteins during neural embryogenesis, and in reactive and neoplastic pathological conditions in the brain (6 –10 ). In recent years, the sensitivity of ISH techniques has increased, using both radioactive and nonradioactive probes to allow the detection of mRNAs that encode various molecules (3 ,5 ,6 ,10 –17 ) and there is increasing use of nonradioactive ISH to identify a number of genes related to vasculogenesis and angiogenesis in the brain (18 –24 ).