Abstract
This chapter focuses on apoptosis which is characterized by the activation or repression of a series of components in the signaling of the cell, and discusses the techniques used in the laboratory to determine whether a glycosphingolipid (GSL) or a cytokine exerts an apoptotic effect on vascular cells. The methods described are applicable to a wide variety of tissues and cells. Cells undergoing apoptosis display dramatic changes that occur within the nucleus during this process. This change is assessed using electron, light and fluorescent microscopic techniques. The DNA-laddering assay is the most common assay used to determine apoptosis in cultured cells as well as in tissues. The detection of internucleosomal DNA fragmentation by standard agarose gel electrophoresis is an important biochemical hallmark of apoptosis. The detection of DNA laddering employing the ELISA assay is considered problematic when only a few cells in the population have undergone apoptosis. The chapter also discusses the analysis of cell death by flow and laser-scanning cytometry and the role of sphingoglycolipids in intracellular signaling/apoptosis.
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