X-ray diffraction patterns from microtubules and neurofilaments in axoplasm

Freshly extracted axoplasm from giant axons of the marine fan worm Myxicola infundibulum and the squid Loligo can be pulled into fibres that contain highly oriented cytoskeletal elements suitable for X-ray diffraction. A major advantage of studying axoplasmic components by this technique is that it allows essentially native structures and their interactions to be examined. We describe here the analyses of the X-ray diffraction patterns. We show that in Myxicola the pattern can be explained by diffraction from both neurofilaments and microtubules, whilst in Loligo the pattern arises solely from microtubules. At low resolution, X-ray patterns obtained from dehydrated axoplasmic microtubules resemble strongly the Fourier transforms generated from electron micrographs of negatively stained specimens. Hydration of axoplasmic fibres produced reversible changes in the X-ray pattern intensities, although the layer-line positions were unaltered. On the 4 nm layer-line, the intensity of the J3 reflection was dramatically reduced on hydration, though its position was unchanged. Hydration also affected the J10/J16 reflections, which increased in intensity, though here again the positions of the peaks were little altered. The X-ray patterns from our hydrated fibres resemble those produced by others from fibres of purified microtubules, though in our patterns contrast is generated towards the centre of the wall. We interpret our findings in the light of current ideas about microtubule structure as determined by X-ray diffraction and electron microscope techniques.