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Ionized calcium concentrations in squid axons

R Dipolo, J Requena, F J Brinley

    The Journal of General Physiology
    |April 1, 1976
    PubMed
    Summary
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    This study measured ionized calcium in squid axons using aequorin and arsenazo dyes, revealing intracellular calcium levels and buffering capacities. Findings highlight calcium

    Area of Science:

    • Neuroscience
    • Biochemistry
    • Cell Biology

    Background:

    • Intracellular calcium regulation is crucial for neuronal function.
    • Accurate measurement of ionized calcium ([Ca]) in axons is essential for understanding cellular processes.

    Purpose of the Study:

    • To determine the concentration of ionized calcium ([Ca]) within squid axons.
    • To investigate the buffering mechanisms of calcium within the axoplasm.
    • To assess the impact of ionic environment on intracellular calcium levels.

    Main Methods:

    • Utilized aequorin luminescence and arsenazo dye spectrophotometry for [Ca] measurement.
    • Employed CaEGTA/EGTA buffers for calibration of [Ca] measurements.
    • Manipulated external ionic composition (Na+, Li+, Choline) and stimulated axons to observe [Ca] dynamics.

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    Main Results:

    • Ionized calcium ([Ca]) in squid axons was determined to be approximately 20-50 nM.
    • Axonal calcium buffering was found to be substantial, limiting rapid [Ca] changes during stimulation.
    • External sodium ions play a significant role in regulating intracellular calcium.

    Conclusions:

    • Squid axons maintain low resting intracellular ionized calcium levels.
    • Significant endogenous calcium buffering systems are present in the axoplasm.
    • Ionic gradients across the axonal membrane influence intracellular calcium homeostasis.