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Related Experiment Videos

Microanalysis of calcium in nerve

F J Brinley, L J Mullins

    Scanning Electron Microscopy
    |January 1, 1980
    PubMed
    Summary

    Researchers analyzed free calcium in squid nerve fibers using aequorin and Arsenazo III. They found resting free calcium levels are low, with organelles like the endoplasmic reticulum buffering most cellular calcium.

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    Ion fluxes in dialyzed squid axons.

    The Journal of general physiology·2009

    Area of Science:

    • Neuroscience
    • Cell Biology
    • Biochemistry

    Background:

    • Intracellular free calcium (Ca2+) is a critical signaling molecule in neurons.
    • Understanding calcium dynamics is essential for comprehending nerve function and dysfunction.
    • Accurate measurement of low nanomolar calcium concentrations is technically challenging.

    Purpose of the Study:

    • To quantify free calcium concentrations in single squid nerve fibers.
    • To identify the primary cellular calcium buffering mechanisms in axoplasm.
    • To explore the role of organelles in maintaining resting calcium levels.

    Main Methods:

    • Utilized the photoprotein aequorin for calcium detection.
    • Employed Arsenazo III, a spectral dye sensitive to calcium binding.
    • Applied these analytical methods to single, isolated squid nerve fibers and axoplasm.

    Main Results:

    • Established that free calcium in squid nerve fibers can be accurately measured in the 20–60 nM range.
    • Determined the resting free calcium concentration in fresh squid axoplasm to be 20–50 nM.
    • Observed that approximately 0.1% of total cellular calcium is ionized, with the remainder buffered by organelles.

    Conclusions:

    • Smooth endoplasmic reticulum and mitochondria are significant calcium buffers in squid axoplasm.
    • Organelles play a crucial role in maintaining low, physiologically relevant levels of ionized calcium.
    • These findings provide insight into neuronal calcium homeostasis and signaling.

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