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D S Gilbert

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Neurofilaments.

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Animals

Axoplasm architecture and physical properties as seen in the Myxicola giant axon.

A technique is described for extracting axoplasm from the giant axon of a marine worm, Myxicola infundibulum. The operation can be completed in 10 sec. 2. Axoplasm is pulled from the axon of a living worm as a long, clear cylinder, up to 35 cm long and 70 mg wet weight. The worm regenerates a new giant axon in about 4 months. 3. Myxicola axoplasm is a gel, 87% water, held together by protein neurofilaments. It contains small amounts of mitochondria and vesicles, but no detectable microtubules. 4. The internal structure of the gel is superficially similar to that of yarn. Closer inspection with light and electron microscopy, and X-ray diffraction, show it to be organized in a hierarchy of helical forms. Squid giant axons have a similar structure. 5. Initial estimates of the bulk physical properties of extracted Myxicola axoplasm give: breaking strength, 1400 g/cm2; specific gravity, 1-05 g/cm3; birefringence, 1-6 X 10(-4); index of refraction, 1-351; resistivity, 57 omega cm. These average values are shown to be compatible with the observed structure and composition. 6. Despite its mechanical strength, the axoplasm gel is so hydrated that Na+, K+ and homarine diffuse through it at rates approaching those in free solution. Fewer than about 5% of each of these ions are tightly bound to the gel. 7. It is argued that (a) the structure and physical properties of Myxicola axoplasm are representative of those in other axons, (b) the compound helix architecture results from twist of parallel, cross-linked fibrous proteins, and (c) this sturcture serves as a flexible internal skeleton for nerve cell processes.

Animals

Axoplasm chemical composition in Myxicola and solubility properties of its structural proteins.

The chemical composition of axoplasm extracted from the giant axon of Myxicola infundibulum has been analysed, and some of the factors which disperse its gel structure have been identified. 2. The axoplasm contains about 3-6% protein, and 0-12% lipid. It is isosmotic with sea water and has a pH near 7-0. 3. Inorganic ions in extracted axoplasm include: Na+, 13m-mole/kg wet wtl; K+, 280; Cl-, 24; Ca2+, 0-3; Mg2+, 3. 4. Free organic ions in axoplasm include: gly, 180 m-mole/kg wet st.; cysteic acid, 120; asp, 75; glu, 10; ala, 7; tau, 5; thr, 2; gln and ser, trace; homarine, 63; isethionate, 0. 5. The gel structure is dispersed by solutions containing 1--10 mM-Ca2+, because this ion activates an endogenous protease. The gel can also be dispersed without proteilysis by solutions containing 0-5 M-KCl, or 0-5 M guanidine hydrochloride, or 3-5 M urea, all of which break down neurofilaments. 6. It is argued that many aspects of the composition and dispersal properties of Myxicola axoplasm are similar to those in other axons.

Amino Acids