Brady Apoptosis Methods and Protocols

Erscheinungsjahr 2008
ISBN: 978-1-59259-812-0
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 282, 304 Seiten, eBook

Reihe: Methods in Molecular Biology

The most fundamental question facing each and every cell within an org- ism is to survive or to die. Cell death is required for normal function; some estimates suggest that as many as one million cells undergo cell death every second in the adult human body. Almost all cells undergoing physiological, or programmed, cell death, independent of cell type, manifest a stereotypic p- tern of morphological changes termed apoptosis. Typically, apoptotic cells d- play shrinkage, membrane blebbing, chromatin condensation, and nuclear fragmentation. The integrity of the cell membrane is not lost during apoptosis and so avoids eliciting the inflammatory response that would have been caused by the spillage of the cell’s contents. This is quite in contrast to the loss of cell contents typical of necrosis. The caspases, the family of intracellular cysteine proteases associated with apoptosis, are responsible for the stereotypical m- phological changes. Caspases cleave various substrate proteins that act on DNA fragmentation, nuclear envelope integrity, the cytoskeleton, and cell volume regulation. Apoptotic cells are cleared in vivo by the process of phagocytosis, in which specific “phagocytes” move to the site of apoptosis, engulf the dying cells and digest them. Apoptosis has a central role in many physiological processes, for example, in the immune system. Autoreactive cells are deleted via apoptosis to prevent autoimmunity. At the end of an immune response, activated lymphocytes are removed to maintain homeostasis within the immune system.

Zielgruppe

Research

Autoren/Hrsg.

Brady, Hugh J. M.

Inhaltsverzeichnis

Measurement of Apoptosis by DNA Fragmentation.- Measurement of Caspase Activity in Cells Undergoing Apoptosis.- Flow Cytometry-Based Methods for Apoptosis Detection in Lymphoid Cells.- Methods of Analyzing Chromatin Changes Accompanying Apoptosis of Target Cells in Killer Cell Assays.- Detecting and Quantifying Apoptosis in Tissue Sections.- Immunoassay for Single-Stranded DNA in Apoptotic Cells.- Methods to Measure Membrane Potential and Permeability Transition in the Mitochondria During Apoptosis.- Measurement of Changes in Intracellular Calcium During Apoptosis.- Measurement of Changes in Apoptosis and Cell Cycle Regulatory Kinase Cdk2.- Assays to Measure Stress-Activated MAPK Activity.- Methods for Culturing Primary Sympathetic Neurons and for Determining Neuronal Viability.- Immunocytochemical Techniques for Studying Apoptosis in Primary Sympathetic Neurons.- Measuring Programmed Cell Death in Plants.- Detection of Apoptosis in Drosophila.- Measurement of Apoptotic Cell Clearance In Vitro.- Yeast Two-Hybrid Screening for Proteins Interacting With the Anti-Apoptotic Protein A20.- The Yeast Three-Hybrid System As a Tool to Study Caspases.- Cloning of Apoptosis-Related Genes by Representational Difference Analysis of cDNA.- Identification of Apoptosis Regulatory Genes Using Insertional Mutagenesis.