Detection and Quantification of Apoptosis in the Vasculature

The integral role of apoptosis in the pathogenesis of cardiovascular diseases has been extensively studied and characterized in recent years. The study of cell death in the vasculature has significantly contributed to our knowledge of vascular disease pathology and has played a role in identifying potential therapeutic strategies for these diseases. This chapter describes a number of standard, widely used protocols for detecting and quantifying apoptosis in vessel wall cells and tissue. These techniques include terminal deoxynucleotidyl transferase dUTP nick-end labeling staining for DNA fragmentation, Hoechst staining for chromatin condensation, Annexin V staining, labeling for phosphatidylserine externalization, Western blot assessment of caspase cleavage, immunofluorescence detection of caspase activation, assessment of mitochondrial membrane depolarization and cytochrome c release, and a splenocyte assay for quantifying susceptibility to immune cell-mediated apoptosis.