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Related Experiment Videos

Reversible structure transition in gap junction under Ca++ control seen by high-resolution electron microscopy.

N G Wrigley, E Brown, R K Chillingworth

    Biophysical Journal
    |January 1, 1984
    PubMed
    Summary

    Rat liver gap junctions undergo structural changes in response to calcium, potentially controlling cell communication. These alterations are reversible but variable, with a connective matrix observed between connexons.

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    Area of Science:

    • Cell biology
    • Biophysics
    • Structural biology

    Background:

    • Gap junctions facilitate direct cell-to-cell communication through communicating channels.
    • These channels are formed by trans-membrane protein assemblies called connexons.
    • Calcium ions are known to influence gap junction permeability and uncoupling.

    Purpose of the Study:

    • To investigate the structural alterations of rat liver gap junctions.
    • To understand the role of calcium in gap junction structural changes and permeability control.
    • To visualize the fine structure of connexons and their arrangement.

    Main Methods:

    • High-resolution, low-dose electron microscopy of deoxycholate-extracted rat liver gap junctions.
    • Imaging in the absence of negative stain to avoid ionic complications.

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  • Digital Fourier filtering and reconstruction for low-contrast images.
  • Negative stain electron microscopy at neutral pH.

    • Main Results:

    • Observed structural alterations in gap junctions upon exposure to calcium (Ca++) down to 10(-4) M.
    • Demonstrated reversible alterations with EGTA, though with considerable variability.
    • Identified a fine connective matrix between connexons using negative stain, potentially explaining lattice invariance during structural transitions.
    • Recovered repeating detail to nearly 1.0 nm resolution in negative stain experiments.

    Conclusions:

    • Calcium induces reversible structural changes in rat liver gap junctions, suggesting a mechanism for permeability control.
    • A connective matrix between connexons may maintain lattice structure during these transitions.
    • Low-dose electron microscopy, despite challenges, provides insights into gap junction dynamics.