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Related Experiment Videos

Ascorbic acid and catecholamine release from digitonin-treated chromaffin cells.

K Morita, M Levine, E Heldman

    The Journal of Biological Chemistry
    |December 5, 1985
    PubMed
    Summary

    Ascorbic acid in bovine adrenal chromaffin cells is released from the cytoplasm, not vesicles, upon cell permeabilization. This finding suggests a non-vesicular storage mechanism for ascorbic acid in these cells.

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    Area of Science:

    • Cell Biology
    • Neuroscience
    • Biochemistry

    Background:

    • Bovine adrenal chromaffin cells are crucial for studying catecholamine and ascorbic acid storage and release.
    • Understanding the subcellular localization of these molecules is key to elucidating their physiological roles.

    Purpose of the Study:

    • To investigate the subcellular localization of ascorbic acid within cultured bovine adrenal chromaffin cells.
    • To determine if ascorbic acid is stored in a vesicular or cytoplasmic pool.

    Main Methods:

    • Cells were permeabilized using digitonin, a steroid glycoside.
    • Catecholamine and [14C]ascorbic acid release were measured under varying calcium concentrations and temperatures.
    • The release of dimethylepinephrine, a vesicular-excluded analog, was also assessed.

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    Main Results:

    • Catecholamine release was calcium-dependent, while [14C]ascorbic acid release was calcium-independent.
    • Ascorbic acid release sensitivity to digitonin matched calcium-dependent catecholamine release.
    • Dimethylepinephrine release kinetics were similar to ascorbic acid release, independent of calcium.

    Conclusions:

    • Ascorbic acid appears to be released directly from the cell cytoplasm upon plasma membrane permeabilization.
    • Newly accumulated ascorbic acid in chromaffin cells is likely localized to a free cytoplasmic pool, not a vesicular compartment.