By Myrtle A. Davis
Dr. Myrtle A. Davis has assembled a panel of state of the art scientists to explain their most sensible equipment for detecting, illuminating, and quantifying apoptotic mechanisms in a manner that's necessary for the layout of toxicology and pharmacology experiences. those state of the art thoughts comprise stream cytometric, fluorometric, and laser scanning equipment for quantifying and characterizing apoptosis, in addition to protocols for using DNA microarray know-how, excessive throughput monitors, and ELISA. Immunocytochemical tools for measuring biochemical and molecular endpoints in tissue sections may be hugely necessary for these accomplishing reviews in complete animal types rather than mobilephone tradition structures.
Read Online or Download Apoptosis Methods in Pharmacology and Toxicology: Approaches to Measurement and Quantification (Methods in Pharmacology and Toxicology) PDF
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Additional info for Apoptosis Methods in Pharmacology and Toxicology: Approaches to Measurement and Quantification (Methods in Pharmacology and Toxicology)
These methods were initially designed to be used by flow cytometry. The drawback of flow cytometric methods stems from the fact that identification of apoptotic or necrotic cells relies on a single attribute that is assumed to represent a characteristic feature (hallmark) of apoptosis or necrosis. , refs. 4–6). The apoptotic attribute may also be absent when apoptosis is induced by the agents that directly or indirectly suppress the appearance of this attribute. The characteristic changes in cell morphology (7), therefore, still remain the gold standard for recognition of apoptotic cell death.
The inner mitochondrial membrane is uniquely suited to act as a proton-diffusion barrier and the F0/F1-ATPase couples diffusion of protons down this gradient (6^µ) to the generation of ATP. Alterations in 6^µ, (usually decreased 6^µ, although some investigators have identified early hyperpolarization) is a common alteration in apoptosis. A number of lipophilic cationic dyes selectively stain mitochondria in proportion to 6^ µ . Some of the more commonly used probes include: rhodamine 123, tetramethylrosamine, MitoTrackerTM rosamine derivatives, DiOC6 (3), and JC-1 (20).
Chem. 276(11), 8079–8086. Goldstein, J. , Waterhouse, N. , Evan, G. , and Green, D. R. (2000) The coordinate release of cytochrome c during apoptosis is rapid, complete and kinetically invariant. Nat. Cell Biol. 2(3), 156–162. , Pazienza, T. , and Muscatello, U. (1997) Effect of cardiolipin on functional properties of isolated rat liver mitochondria. Biochem. Mol. Biol. Int. 41(3), 469–480. , and Wang, X. (2000) Cardiolipin provides specificity for targeting of tBid to mitochondria. Nat. Cell Biol.