Removal of the Schwann sheath from the giant nerve fiber of the squid: an electron-microscopic study of the axolemma and associated axoplasmic structures.

The axolemma is associated structurally and functionally with the axoplasm, forming an axolemma-ectoplasm complex. To study the structure of this complex, a new technique was developed for removing the Schwann sheath from a portion of the giant nerve fiber. An isolated fiber was treated, without loss of excitability, with trypsin dissolved in natural seawater. Next, the fiber was treated with a mild fixative and then was placed in a hypertonic solution of sucrose in seawater. The elevated sheath was transected and everted, thus exposing the surface of the axon. Desheathed axons were examined by scanning and transmission electron microscopy. The surface of the axon has a ridge-and-groove pattern, reflecting an underlying helical arrangement of filaments which bundle and unbundle. Both left and right axons of the squid possess right-handed helical twists with a tilt angle of 10 degrees. Hemispherical protuberances about 1.5 microns at their base are observed along the ridges. Thin sections of the desheathed axons reveal that the desheathing procedure leaves the axolemma intact. Desheathed axons display electron-dense bodies associated with the axolemma and with the filaments of the ectoplasm similar to the dense bodies observed in whole fibers fixed in the presence of 10 mM Co(II) ions. Axons perfused for 40 min with a solution containing 2 mM Co(II) ions retain their excitability and display a smooth inner ectoplasmic face. A portion of the axolemma, together with adhering ectoplasm, was removed from desheathed axons, mounted between folding double grids, stained, and critical-point dried. Through this novel method a network of 10 nm filaments spaced 40 nm apart and cross-linked by filaments 5 to 7 nm in diameter was demonstrated.