Intracellular levels and distribution of Ca2+ in digitonin-permeabilized cells.
Digitonin and other saponins can be used to selectively permeabilize the plasma membrane of a wide variety of cells without significantly affecting the gross structure and function of Ca2+-sequestering organelles such as mitochondria and endoplasmic reticulum. These characteristics have allowed digitonin to be used in the determination of the intracellular levels and distribution of Ca2+, as well as the measurement of Ca2+ fluxes by organelles "in situ". Studies conducted with several different types of digitonin-permeabilized cells indicate that the endoplasmic reticulum functions as a high affinity and low-capacity intracellular Ca2+ buffer, whereas mitochondria operate as a relatively low affinity but high capacity Ca2+ buffering system. However, recent findings suggest that mitochondria have a comparable affinity for net Ca2+ uptake in the presence of physiological concentrations of polyamines. The use of permeabilized cells has also been important in the identification of the endoplasmic reticulum as a site at which the recently discovered second messenger inositol trisphosphate acts to bring about an increase in the cytosolic free Ca2+ concentration. Thus, the selective permeabilization of cells with digitonin and its analogues has been a powerful yet simple tool in the study of intracellular Ca2+ homeostasis.
D000818:Animals; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism*; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism*; D004072:Digitonin / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000378:metabolism; D008563:Membrane Lipids; D008928:Mitochondria / Q000378:metabolism; D051381:Rats; D012503:Saponins / Q000494:pharmacology; D012995:Solubility; D013096:Spermine / Q000378:metabolism
D008563:Membrane Lipids; D012503:Saponins; D013096:Spermine; D004072:Digitonin; D002118:Calcium