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Atomic force microscopy and dissection of gap junctions.

J H Hoh1, R Lal, S A John

  • 1Division of Biology, California Institute of Technology, Pasedena 91125.

Science (New York, N.Y.)
|September 20, 1991
PubMed
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Atomic force microscopy revealed hepatic gap junction structure. Force dissection exposed hexagonal hemichannel arrays, showing a 9.1 nm spacing, consistent with electron microscopy findings.

Area of Science:

  • Cell biology
  • Biophysics
  • Microscopy

Background:

  • Hepatic gap junctions are crucial for intercellular communication in the liver.
  • Previous structural studies relied on electron microscopy, limiting dynamic analysis.
  • Understanding gap junction ultrastructure is key to deciphering liver function and disease.

Purpose of the Study:

  • To investigate the detailed structure of isolated hepatic gap junctions using atomic force microscopy (AFM).
  • To explore the mechanical properties and dissection capabilities of AFM on gap junction membranes.
  • To visualize the arrangement of hemichannels within the gap junction structure.

Main Methods:

  • Atomic force microscopy (AFM) was employed to image isolated hepatic gap junctions in phosphate-buffered saline (PBS).

Related Experiment Videos

  • Force titration experiments were performed to "dissect" the top membrane layer.
  • Samples were treated with trypsin and fixed with glutaraldehyde prior to AFM analysis.

    • Main Results:

    • The thickness of hepatic gap junctions was measured to be approximately 14.4 nm, aligning with electron microscopy (EM) data.
    • Force dissection successfully removed the top membrane, exposing the extracellular domains of the bottom membrane.
    • A hexagonal array of gap junction hemichannels was visualized, exhibiting a center-to-center spacing of 9.1 nm.

    Conclusions:

    • AFM provides high-resolution structural insights into hepatic gap junctions.
    • Force dissection is a viable technique for revealing the underlying hemichannel organization.
    • The observed hexagonal array and spacing support current models of gap junction assembly and function.